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Sugars:
Monosachharides simple sugars (can not be broken down to simpler sugars under mild conditions)
Monosachharides .
2 :
- aldose ( , ) - ketose ( , )
Polysaccharides
Storage Glycogen The glucose storage device in animals (present in all cells)
Glycogen constitutes up to 10% of liver mass and 1-2% of muscle mass Glycogen is stored energy for the organism Only difference from starch: number of branches Alpha(1,6) branches every 8-12 residues Like amylopectin, glycogen gives a red-violet color with iodine
Starch
Storage
Amylose is a linear polymer of glucose and poor soluble in water. Forms helical structure Is a storage molecule in plants Why good for storage? Glucose monomers diffuse in the cell. Too much glucose would cause osmotic pressure. Therefore, cells arrange the glucose in long polymers. When glucose is required the enzymes remove monomers from the ends.
Amylopectin branched polymer of glucose which provide a mechanism for quickly releasing (or storing) of glucose units for (or from) metabolism
Starch
In amylose - iodine fits into the helices to produce a blue color, a way of staining cells when testing for starch.
Amylopectin - Does not form helices because branches every 24-30 residues and need 6 residues per turn in a helix.
Storage Glycogen
Structure: Similar to amylopectin but more branches, that allow rapid mobilization of glucose when required.
Glycogen
Glycogen is a polymer of glucose residues linked by a(14) glycosidic bonds, mainly a(16) glycosidic bonds, at branch points. Glycogen chains & branches are longer than shown.
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Polysaccharides
Structure
Plants rely on polysaccharides for structure. Do not synthesize keratin and collagen.
Cellulose Cellulose consisting of a linear chain of several hundred to over ten thousand linked glucose units Half the carbon in the biosphere is in the form of cellulose Cellulose is the most abundant natural polymer on earth Provides strength for plants and trees But can also be soft in cotton Plant cell wall
Cellulose
Structure
The 1-4 link causes a fully extended linear polymers and can form hydrogen bond with polymers that are parallel to it Animal enzymes do not have enzymes that cleave the 1-4 bond. Therefore humans can not utilize the vast cellulose around them. Cow intestines have symbiotic bacteria that can digest cellulose. Slow process.
Converting cellulose from energy crops into biofules is under investigation as an alternative fuel source .
Structure
Cellulose
Form sheets
Catalytic site
Binding site
Substrate
Enzyme
+ Enzyme
ES Substrate Complex
The product is finally made and the enzyme is ready for another substrate. EP E + P
- Ea
- , , . , . . , .
- . , , . .
ATP
Cells need energy in the form of ATP. How does the cell generate ATP?
Metabolism The process by which living organisms acquire and use free energy to carry out their functions: Catabolism (degradation) Breakdown of nutrients to get components and/or energy Anabolism (biosynthesis) Biomolecules are synthesized from simpler components
DNA Nucleotide
Phosphate Group
O O=P-O O
CH2
O
N
C
Sugar (deoxyribose)
P
5 4 3 2 1
O O
1
3
4
P
5
T
O
A
O
BASE-PAIRINGS
H-bonds
PCR
ATP- .ADP
. ( ). , .
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Glycolysis
O CH3 C
Ethanol
CO2
Glucose enters the cell through a specific carrier transports glucose into the cytosol
The glycolytic enzymes are located in the cytosol. They are loosely associated with cell structures and membranes.
Glycolysis has 2 phases: Converts glucose to two G-3-P with energy input
* The kinase activity of the enzyme: transfers a phosphoryl group between ATP and a metabolite
Glc
* Glu-6-P can regulate hexokinase activity by inhibition: it can compete for the active site and also by allosteric interactions at another site of HK.
* Because glucose and fructose are cyclic - the isomerization requires ring opening. * The intermediate is called an enediol
* This reaction is reversible
* Isomerization of an aldose to a ketose * Why is this step needed? Carbonyl oxygen (C=O) is transferred from C1 to C2, and the OH on C1 is accessible for the next phosphorylation. (Would be hard to do on glucose). This also prepares the molecule for cleavage in step 4.
* 2nd ATP investment. Phosphorylated on two carbons. * Spontaneous and irreversible reaction. * Reaction is similar to HK. * PFK is the rate-limiting enzyme in glycolysis. * Enzyme is highly regulated will be discussed.
4 5 6 1 2
Preparatory stage:
* Since only GAP continues in glycolysis, this step is required so all glucose carbons are utilized.
* Since GAP is low in cells (since GAP is used up in the downstream reactions) the reaction is actually exerogenic
Keq = [GAP]/[DHAP] = 4.73 x 10-2 [DHAP]>>[GAP] but because GAP is utilized downstream, more DHAP is converted to GAP
* First high energy intermediate is formed acyl phosphate. * The phosphate group has high free energy of hydrolysis: G = -49.4 kJ/mol
* NAD is a co-enzyme that accepts electrons and a H+ from the substrate, and is reduced to NADH.
Nicotinamide Adenine Dinucleotide (NAD) is a coenzyme derived from the vitamin niacin, or B3. Is important in reduction and oxidation reactions. (oxidation = loose electrons; reduction = gains electrons)
NAD+ picks up H2 and briefly becomes NADH2+, then quickly stabilizes by losing H+ and becoming NADH
(1,3-BPG)
* First generation of ATP * At this point the net ATP yield is zero (from 2 GAPs)
* (Called a kinase because transfers a phosphoryl between a metabolite and ATP, although it is the reverse reaction)
Reactions 6 (G 0=+6.7) and reaction 7 (G0= -18.8) are coupled G0 =-12.1 kJ/mol
* Phosphate is shifted from C3 to C2 * Requires magnesium * [3-PG] > [2-PG] so the reaction is actually exergonic in cells
tetramer
* Spontaneous reaction
* Removal of Pi from PEP yields an unstable enol, which spontaneously converts to the keto form of pyruvate * Highly exergonic and drives ATP synthesis * Hydrolysis of the phosphate in Phosphoglycerate will give only a G of 17.6 kJ/mol not enough to drive ATP synthesis But PEP has a larger G of phosphate hydrolysis than ATP. (PEP -61 kJ/mol, ATP -31 kJ/mol)
Glycolysis
Balance sheet for ~P bonds of ATP: 2 ATP expended 4 ATP produced (2 from each of two 3C fragments from glucose) Net production of 2 ~P bonds of ATP per glucose.
The fate of pyruvate and NADH Aerobic or anaerobic: 1) NADH is energy - two possible fates:
* If O2 is available: NADH is re-oxidized in the electron transport pathway, making ATP in oxidative phosphorylation (1NADH=3ATP)
* In anaerobic conditions: NADH is re-oxidized by lactate dehydrogenase (LDH), providing additional NAD+ for more glycolysis
2) Pyruvate is energy - two possible fates: * Aerobic: citric acid cycle * Anaerobic: Animals, bacteria - LDH makes lactate Yeast alcoholic fermentation
NADH: must be changed back to NAD+ for glycolysis to continue (little NAD+ in cells) * Aerobic conditions mitochondrial electron transport NADH oxidized & ATP energy
* Anaerobic conditions: 1. Bacteria dont have mitochondria (no respiratory chain) 2. High rates of glycolysis in animal cells, like skeletal muscle during exercise - there is no more oxygen and high demand for ATP, and not enough energy to support it Pyruvate is converted to lactate, regenerating NAD+ needed for glycolysis, the main source of ATP under anaerobic conditions.
3. Alcoholic fermentation
* Lactate diffuses from tissue to blood and transported to heart and liver, that convert it back to glucose * High lactate in muscle does not cause fatigue, but instead the buildup of acid. As a result pH drops and causes Bohr effect in order to increase oxygen delivery to the tissues.
Lactate dehydrogenase
Tetrameric enzyme, composed of two subunits M and H. Enzyme exists in different tissues as isozyme (different aa sequence of the enzymes that catalyze the same reaction). Two subunits can randomly bind one another, so it can be 5 isozymes: M4, M3H, M2H2, M2H3, H4.Each isozyme is specific for given tissue. Clinical significance: LDH pattern in serum can tell us about pathological condition
Control of glycolysis
Control points: Enzymes that catalyze reactions that are highly exerogenic (irreversible) Allosteric enzymes Large negative free energy changes are found in only: 1) Hexokinase 2) Phosphofructokinase 3) Pyruvate kinase
HK
PFK
PK
( ) = micromolar concentrations
Control of glycolysis
Control of glycolysis
In liver cells there is a hexokinase variant: Glucokinase Has a high Km for glucose active only at high glucose.
This allows liver cells to store glucose as glycogen when blood levels are high. So G6P does not inhibit glucokinase activity (G6P is a substrate for glycogen synthesis).
Control of glycolysis
Glucokinase
When glucose is required in the blood enzyme glucose-6-phosphatase will release glucose to the bloodstream.
Glucokinase
The enzymes Glucokinase & Glucose-6-phosphatase, are both found in liver but not in most other body cells. This allows the liver to control blood glucose levels.
Control of glycolysis
Phosphofructokinase The rate-limiting enzyme in glycolysis
Because: 1) HK has a variant that acts differently in liver 2) Pyruvate kinase is the last step too late for regulation
ATP is both a substrate and an allosteric inhibitor: there are 2 binding sites for ATP in each subunit a substrate site, an inhibitory site.
Citrate is a PFK inhibitor. Activators of PFK: ADP, AMP, fructose-2,6-bisphosphate
Control of glycolysis
Phosphofructokinase
PFK is a tetramer
Each subunit has 2 ATP-binding sites: Substrate site or active site and inhibitory site
At low concentration, ATP binds only at the active site. At high ATP: ATP binds to the inhibitory site (low-affinity regulatory site) and decreases the affinity for its substrate Fructose-6-P
Control of glycolysis
Phosphofructokinase
PFK increases activity when energy status is low PFK decreases activity when energy status is high
At high levels of ATP in a cell the cell does not need any more ATP. Therefore it is logical that the ATP itself inhibits its own generation, and it is more useful for the cell to store glucose as glycogen when ATP is plentiful. The tense conformation of PFK: at high [ATP], has lower affinity for the other substrate, fructose-6-P, and binds ATP at the inhibitory site. Sigmoidal dependence of reaction rate on [fructose-6-P] is seen.
AMP, present at significant levels only when there is extensive ATP hydrolysis, antagonizes effects of high ATP.
Control of glycolysis
Phosphofructokinase
Fructose-6-P
PFK-2 Fructose-2,6-BP cAMP dependent protein kinase ATP - cAMP glucagon FBPase-2
Fructose-2,6-BP
PFK-1
Phosphoprotein phosphatase
Fructose-2,6-BP
PFK-1
Control of glycolysis
Pyruvate kinase
Inhibited by acetyl-CoA: If there is enough fatty acid breakdown, then dont need glycolysis
Activated by AMP
Activated by Fructose-1,6,-Bisphosphate (product of PFK1): This ensures that if this product is made that it will pass through the full pathway and that intermediates will not accumulate
Dietary regulation of the enzyme level (after reach carbohydrate meal enzyme level increases up to 10 folds)
cAMP-dependent protein kinase phosphorylates pyruvate kinase and inhibits it.
Thus glucagon-induced production of cyclic-AMP results in inhibition of glycolysis in liver, by slowing of both pyruvate kinase and phosphofructokinase
Glycogen and starch, polymeric storage forms of glucose, enter glycolysis in a two-step process: 1) Phosphorolytic cleavage of a glucose residue from an end of the polymer, forming glucose 1phosphate, is catalyzed by glycogen phosphorylase or starch phosphorylase. 2) Phosphoglucomutase then converts the glucose 1-phosphate to glucose 6-phosphate, which can enter glycolysis.
Feeder pathways for glycolysis Ingested polysaccharides and disaccharides: Disaccharides must be hydrolyzed to monosaccharides before entering cells. Intestinal disaccharides and dextrins are hydrolyzed by enzymes attached to the outer surface of the intestinal epithelial cells:
Feeder pathways for glycolysis D-Galactose is formed mainly by hydrolysis of lactate (milk). D-galactose is converted in a number of reactions to glucose-6-phosphate
Galactosemia a genetic disorders in galactose usage. As a result galactose is accumulated and causes eye cataracts, decrease in mental development and other problems. The disease is found mainly in infants.
Lactase is an enzyme that catalyses lactose breakdown to glucose and galactose. In some individuals enzyme disappears from the small intestine cells at the age 4-6. This condition is called lactose intolerance
Animations http://programs.northlandcollege.edu/biology/Biology1111/animations/glycolysis.html
http://www.iubmb-nicholson.org/swf/glycolysis.swf
http://www.maxanim.com/biochemistry/How%20Glycolysis%20Work/How%20Glycoly sis%20Work.htm
Oxidation of pyruvate to acetyl-coA is a multiple step process which involves: - pyruvate decarboxilation - NAD+ reduction - pyruvate oxidation - pyruvate activation by coenzyme A
Pyruvate dehydrogenase
Pyruvate dehydrogenase complex is composed of three enzymes: Pyruvate dehydrogenase (E1) Dihydrolipoamide transacetylase (E2) Dihydrolipoamide dehydrogenase (E3) and five coenzymes Thiamine Pyrophosphate (TPP) Lipoic Acid-Lipoamide FAD NAD+ CoASH
Pyruvate dehydrogenase
In eukaryotes the mitochondrial PDH has: 20 E2 trimers 30 E1 heterotetramers 12 E3 dimers
This is an example in which intermediate products are passed from one enzyme to another. If the enzymes depend only on diffusion then this is very efficient. Also, they are passed to each other and side reactions are minimized. The enzymes can be controlled together.
Five coenzymes are used - TPP, CoASH, Lipoic acid, NAD+, FAD There are 5 reactions, the net stoichiometry is: Pyruvate + CoA + NAD+ acetyl-CoA + CO2 + NADH
Thiazole ring
Pyrimidine ring
- TPP -keto
Pyruvate dehydrogenase
Pyruvate dehydrogenase
Pyruvate dehydrogenase multienzyme complex (from Bacillus) rendered from 3D data with lipoyl domains added by hand. The lipoyl arms move the intermediates between the enzymes.
Pyruvate dehydrogenase
Pyruvate dehydrogenase multienzyme complex (from Bacillus) rendered from 3D data with lipoyl domains added by hand. The lipoyl arms move the intermediates between the enzymes.
Pyruvate dehydrogenase
glucose-6-P Glycolysis pyruvate fatty acids acetyl CoA oxaloacetate Krebs Cycle ketone bodies cholesterol citrate
Acetyl CoA functions as: input to Krebs Cycle, where the acetate moiety is further degraded to CO2. donor of acetate for synthesis of fatty acids, ketone bodies, & cholesterol. Since they are no other pathways for making acetyl-coA from pyruvate, this pathway must be regulated
Pyruvate dehydrogenase Regulation of Pyruvate Dehydrogenase Complex by product inhibition: 1) Product inhibition by NADH & acetyl CoA: NADH competes with NAD+ for binding to E3. Acetyl CoA competes with CoA for binding to E2. NADH and acetyl-CoA are products of fatty acid oxidation, so when lipids are available the cell will preserve stores of carbohydrates.
Pyruvate dehydrogenase
Regulation of Pyruvate Dehydrogenase Complex by phosphorylation/dephosphorylation (only in eukaryotes): 2) Phosphorylation of E1 inhibits the complex: A family of Pyruvate Dehydrogenase Kinases catalyze phosphorylation of E1, inhibiting the complex. These kinases are activated by NADH & acetyl CoA (another way the major products inhibit the complex). Amounts of these Kinases increase in muscle mitochondria during starvation, effectively blocking catabolism of glucose (glycolysis), since glucose is needed by the brain.
A Ca++-sensitive isoform of the phosphatase that removes Pi from E1 , is expressed in muscle cells. Increased cytosolic Ca++ can lead to Ca++ uptake by mitochondria, & activation of Pyruvate Dehydrogenase. Since muscle contraction involves Ca, metabolism may be stimulated during exercise. Pyruvate Dehydrogenase Kinase and Phosphatase enzymes are unique. They bind specifically to Pyruvate Dehydrogenase within the mitochondrial matrix.
Krebs cycle
(Hans Krebs, Nobel prize 1953)
Cycle:
a multistep cycle in which the first step is regenerated in the last step the cycle can oxidize an unlimited number of acetyl groups
In eukaryotes all the enzyme of the cycle are in the mitochondria. This requires either production of metabolites in the mitochondria or transport out.
Localization of Krebs Cycle: Glycolysis occurs in the cytosol of cells. Pyruvate enters the mitochondrion to be metabolized further.
Mitochondrial Compartments:
The matrix contains Pyruvate Dehydrogenase, enzymes of Krebs Cycle, and other pathways, e.g., fatty acid oxidation & amino acid metabolism. The outer membrane contains large VDAC channels. The outer membrane is leaky to ions & small molecules.
Mitochondrial compartments
Inner membrane infoldings, called cristae, contain constituents of the respiratory chain & ATP Synthase. The inner membrane is the major permeability barrier. It contains various transport catalysts, including a carrier protein that allows pyruvate to enter the matrix.
Other names: Phosphogluconate Pathway Hexose Monophosphate Shunt The linear part of the pathway carries out oxidation and decarboxylation of the 6-C sugar glucose-6-P, producing the 5-C sugar ribulose-5-P. Takes place in the cytosol
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The energy currency in cells is ATP. But cells also use: reducing power. The Pentose Phosphate Pathway provides:
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Erythrocytes and the cells of the lens and cornea are directly exposed to oxygen and thus to the damaging free radicals generated by oxygen.
By maintaining a reducing atmosphere (a high ratio of NADPH to NADP and a high ratio of reduced to oxidized glutathione), they can prevent or undo oxidative damage to proteins, lipids, and other sensitive molecules.
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Pentose Phosphate Pathway NADPH and NADH are different molecules: Oxidizing enzymes use NADH (catabolic) Reducing enzymes use NADPH (anabolic)
Occurs mainly in tissues that synthesize fatty acids and steroids: cytoplasm of liver, adipose tissue
Reduction of NADP+ (as with NAD+) involves transfer of 2 e- and 1 H+ to the nicotinamide moiety. This difference between NAD and NADP has little to do with redox activity, but is recognized by substrate-binding sites of enzymes. It is a mechanism for separation of catabolic and synthetic pathways.
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Oxidative phase
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This reaction occurs without the enzyme. The enzyme increases the rate of hydrolysis.
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Regulation of Glucose-6-phosphate Dehydrogenase: Glucose-6-phosphate Dehydrogenase is the committed step of the Pentose Phosphate Pathway. This enzyme is regulated by availability of the substrate NADP+. As NADPH is utilized in reductive synthetic pathways, the increasing concentration of NADP+ stimulates the Pentose Phosphate Pathway, to replenish NADPH. NADPH strongly inhibits the enzyme.
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Ribose-5-P is the precursor of nucleotides. If the cell makes too much Ribulose-5-P then it is converted to glycolysis (converted to F-6-P and GAP, that come next)
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2) Ribulose-5-P epimerase
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Pentose Phosphate Pathway We now have C-5 sugars, but will get C-6 sugars in the end of PPP. How is this done? Transketolase: catalyzes the transfer of C-2 units
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From step 5
Previous step
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In the Pentose Phosphate Pathway three C-5 sugars are converted to two C-6 and one C-3 sugars.
C5 + C5 C3 + C7 C3 + C7 C6 + C4
(Transketolase) (Transaldolase)
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Glucose-6-phosphate may be regenerated from either the: 3-C glyceraldehyde-3-phosphate or the 6-C fructose-6-phosphate
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When in the PPP needed? depends on the metabolic needs of the cell
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When in the PPP needed? depends on the metabolic needs of the cell For nucleotide synthesis: 1) Ribose-5-P is needed for nucleotide synthesis 2) Some NADPH is needed to reduce ribonucleotides to deoxyribonucleotides
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When in the PPP needed? depends on the metabolic needs of the cell
When reducing power is needed for fatty acid or steroid synthesis: Generates a lot of NADPH and G-6-P is regenerated to go back into the PPP, and make more NADPH
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When in the PPP needed? depends on the metabolic needs of the cell When only moderate levels of NADPH are needed: The cycle is used to make some energy. The product of PPP: F-6-P and G-3-P are further used by the glycolytic and citric acid pathways to make ATP
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Glutathione
Glutathione is the most abundant antioxidant in the body. It is a tripeptide, made up of cysteine, glutamic acid, and glycine. Glutathione is special in that it is fat soluble and water soluble, therefore it has anti-oxidant activity throughout the entire body. Being an anti-oxidant, glutathione quenches free radicals, thereby protecting healthy cells from damage. Glutathione also acts as a chelator to bind and escort out of the body heavy metals that cause the free radical damage. Glutathione is an important part of the livers detoxification system so it helps to remove metabolic waste. It also has an important role in transporting amino acids into cells
Erythrocytes use glutathione to eliminate H2O2 and hydroperoxides that can damage hemoglobin or the cell membrane. Hydroperoxides, arising spontaneously in the oxygen-rich environment in red blood cells, react with double bonds in fatty acids of membrane lipids, making membranes leaky.
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Glutathione
Glutathione is present in most cells and protects against oxidative stress, and keeps thiol groups of proteins in a reduced state:
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Regeneration of reduced glutathione requires NADPH produced in the Pentose Phosphate Pathway.
Reduced glutathione is generated by glutathione reductase:
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Defects in the PPP pathway Glucose-6-Phosphate Dehydrogenase deficiency People who have this deficiency (decreased activity of the enzyme) are sensitive to oxidative damage, which can be seen as hemolytic anemia
These people if they take anti-malaria drug primaquine or eat fava beans ),( which stimulate peroxide formation, will need high levels of NADPH that the mutant cell cannot supply. Like in sickle-cell anemia, in this deficiency the erythrocytes are resistant to infection by the malaria parasite.
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Defects in the PPP pathway Thiamine (B1) is part of the coenzyme TPP (thiamin pyrophosphate). It promotes conversion of pyruvate to acetyl CoA by removing one carbon Similar reaction in TCA cycle And in transketolase (PPP)
Wernicke-Korsakoff Syndrome Mental disorder due to thiamine deficiency loss of memory, partial paralysis. Particularly in alcoholics that lack this in their diet. Due to a mutated transketolase that has 10-fold reduced affinity to its cofactor thiamine (TPP), the PPP is slowed down. Normally if TPP levels drop this does not affect the enzyme. But in people with the abnormal enzyme this causes disease.
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Characterized by: loss of sensation in hands and feet muscular weakness advancing paralysis abnormal heart action
Found mainly in people from Asian countries eat white rice where the husk () has been removed that part has most of thiamine.
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The major pathways of fuel metabolism in mammals Most tissues cannot carry out all the following reactions.
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Organ specialization
Brain
High respiration rate: responsible for 20% of body oxygen consumption (2% of body mass) High energy requirement about 15% of total energy Most energy is required for the plasma membrane (Na/K)-ATPase needed for maintaining the membrane potential required for nerve pulses. Glucose is the main fuel of the brain (120 gr/day) . During fasting it uses ketone bodies (= products of breakdown of fatty acids) Since there is not much glycogen storage in the brain it requires a constant supply of glucose. Low glucose half of normal 4-5mM results in brain dysfuction. Lower coma and irreversible damage, death.
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Organ specialization
Muscle Fuels: 1) Glucose from glycogen 2) Fatty acids 3) Ketone bodies During rest the muscle generates glycogen storage. Muscle stores about from total glycogen in the body Glycogen is converted to G-6-P for glycolysis. Muscle cells do not have G-6phosphatase so cannot export glucose. Muscle does not have enzymes for gluconeogenesis. Muscle carbohydrate metabolism serves only the muscle.
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Organ specialization
Muscle
Muscle contraction
Muscle contraction requires ATP. Most ATP in the body comes from TCA and oxidative phosphorylation. At rest the muscles use 30% of body oxygen. During high exertion it gets ATP from: phosphocreatine + ADP creatine + ATP
This is good for 4 seconds, then the muscle shifts to glycolysis of G-6-P. The flux is too much for TCA and ox. phos., therefore G-6-P is degraded to lactate (anaerobic).
Lactate is released to blood and converted in liver to glucose (Cori cycle)
Organ specialization
Heart
A muscle that acts continuously relies only on aerobic metaboilsm and has many mitochondria ( up to 40% of the cytoplasm).
Contains little glycogen and stored lipids and small amounts of creatine phopshate
During resting it uses fatty acids. During heavy work needs glucose and uses up its supply of glycogen. Therefore the heart needs constant supply of oxygen and fuels from the blood.
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Organ specialization
Adipose tissue
This tissue must store or release fatty acids as needed. Found mostly under the skin, abdomen, skeletal muscle. A 70 kg person has 15 kg of fat = 590,000 kJ When intracellular glucose levels are low, the hormone sensitive lipase causes fatty acids to be sent into the blood. When glucose levels are OK, there is resynthesis of triglycerols from fatty acids.
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Organ specialization
Liver The center for keeping levels of circulating fuels, for the brain, muscles and other tissues.
Uptake and release of glucose in response to hormones and blood glucose levels. After carbohydrate dinner: blood glucose is 6mM glucose converted to G-6-P by glucokinase (isoenzyme of hexokinase). More glucose more enzyme activity, so when all tissues are satisfied it will uptake glucose from blood. (Normal blood glucose: 4 to 8 mM (70 to 150 mg/dL)
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Organ specialization
Liver Fates of G-6-P in liver: 1) When blood glucose drops under 5mM is converted to glucose by G-6-phosphatase, and travels by blood to organs. During exercise or fasting, low blood glucose causes pancreas to secrete glucagon that acts on liver cells that breakdown glycogen. 2) Converted to glycogen when there is no demand for glucose. 3) Converted to acetyl-CoA via glycolysis and can go to: TCA / fatty acid generation / phospholipids / cholesterol 4) Degraded by the PPP to generate NADPH required for fatty acid synthesis.
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Organ specialization
Kidney
Filters waste and urea and retains metabolites like glucose. Maintains blood pH by exerting excess H+. It exerts protons like NH4+ (ammonia derived from glutamine or glutamate). The remaining amino acid skeleton: -ketoglutarate can be converted to glucose by the citric acid cycle and then gluconeogenesis. (Only done by liver and kidney), or oxidized by ox. phos. to CO2. Blood In erythrocytes (99% of blood cells) glycolysis is the only pathway since they have no mitochondria and therefore depend of anaerobic glycolysis.
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Organ specialization
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Disturbances of metabolism Diabetes Mellitus Third leading cause of death in the USA.
Problem: 1) Insulin is not secreted in efficient amounts, or 2) Insulin does not efficiently stimulate target cells
Results Blood glucose levels are high = hyperglycemia (spill into urine) but cells are still starved for glucose Other pathways become accelerated: gluconeogenesis, triacylglycerol hydrolysis, fatty acid oxidation, and ketone body formation (ketosis) Ketone bodies are acids the body works hard to control blood pH by removing H+, but also other ions are excreted (Na+, K+ ,H2O) and this causes dehydration, and decrease in blood volume.
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Disturbances of metabolism
Diabetes Mellitus
Type I: Insulin-dependent or juvenile-onset diabetes The pancreas lacks or has defective cells (responsible for insulin secretion) Usually an autoimmune response that destroys the cells, noticed after 80% of cells have been eliminated.
Occurs usually in childhood. Daily injections of insulin and balance diet increase the survival, but still there can be many organ complications (kidney, nerves, cardiovascular, cataract)
Successful treatment: cell transplantation
Pancreatic lesion
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Disturbances of metabolism
Diabetes Mellitus
However, in most obese humans leptin levels increase when body fat rises. The defect is probably not in leptin production but in leptin resistance = decrease in the levels of the leptin receptor in the brain (similar to diabetes type II) Also low response to leptin increases neuropeptide Y which in the brain stimulates appetite. There are also other hormones that are involved in obesity.
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Hormonal control of metabolism In animals coordination of activities between organs is done by hormones. Hormones are synthesized by endocrine glands.
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Hormonal control of metabolism Islets of Langerhans endocrine cell that secrete polypeptides into the blood stream cells insulin, in response to high glucose cells glucagon, in response to low glucose
Pancreatic hormones
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Insulin
Pancreatic cells are most sensitive to blood glucose at levels of 5.5 6.0 mM (normal is 3.6-5.8 mM)
Glucose diffuses into the cells and is sensed by the sensor glucokinase (like liver) In these cells all G6P is converted to: pyruvate citric acid cycle oxidative phospohrylation (No glycogen storage, low PPP, low lactate dehydrogenase)
Mechanism is not known but oxidative phosphorylation levels regulate insulin synthesis and secretion.
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Insulin
Insulin promotes glucose uptake in muscle and adipose tissue and inhibits hepatic glucose production.
The Glut4 system allows the tissues to quickly accumulate glucose after a meal.
Once glucose enters cells it can be used for synthesizing: Glycogen in muscle Triacylglycerols in adipocytes Insulin promotes these activities.
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Insulin
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Insulin In liver cells - insulin does not increase rate of glucose uptake. Instead it:
Results: Liver stores glucose as glycogen and triacylglycerols (instead of making glucose by glycogenolysis and gluconeogenesis)
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Muscle cells do not respond to glucagon dont have a receptor, therefore get the glucose indirectly from the liver
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Adrenal hormones
The adrenal glands release hormones in response to neuronal signals. The cortex synthesizes and secretes steroid hormones The medulla synthesizes epinephrine and nor-epinephrine They are released during stress.
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Epinephrine Liver Epinephrine promotes the release of glucagon from the pancreas in liver it stimulates glycogen breakdown Epinephrine binds to receptors on liver cells (adrenergic receptors) and turns on cAMP signal transduction, leading to glycogenolysis and gluconeogenesis
Muscle
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Glycogen If this much glucose were dissolved in the cytosol of a hepatocyte, its concentration would be about 0.4 M, and will cause osmotic problems in the cell. When stored as a long polymer (glycogen), however, the same mass of glucose has a concentration of only 0.01 M. Glycogen granules are complex aggregates of glycogen and the enzymes that synthesize it and degrade it, as well as the machinery for regulating these enzymes.
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Glycogen Glucose is essential for brain and red blood cells, that depend entirely on glucose. When glucose is required one source is glycogen. Glycogen is stored mainly in liver & skeletal muscle. 10% of the weight of liver and 1% to 2% of the weight of muscle. Glycogen from diet meat. Glycogen storage is way for higher organisms to protect themselves from fuel shortage.
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Glycogen
Why does the body store glucose as glycogen, when it could use fat that is more abundant in the body? 1( Muscles cant use fat as rapidly as they can glycogen 2) Fatty acids (from fat) cannot be used anaerobically 3) Animals cannot turn fatty acids into glucose
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Glycogenolysis = glycogen breakdown Glucose is removed from ends of branches - two possible reactions: Hydrolysis: usually for digested glycogen (food). In this reaction elements of water are added to glycosidic bond. Hydrolysis yields glucose that can be absorbed in blood. Phosphorolysis: usually for stored glycogen. In this reaction elements of phosphoric acid are added to glycosidic bond. The advantage is a sugar phosphate that can be utilized directly in the glucose pathways within the cells.
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Glycogenolysis
1. Glycogen phosphorylase
Pyridoxal phosphate (PLP), a derivative of vitamin B6, serves as prosthetic group for Glycogen Phosphorylase.
Lys
In contrast to the role of this cofactor in other enzymes, the phosphate of PLP is involved in the catalytic activity of the enzyme.
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PLP
A class of drugs developed for treating the hyperglycemia of diabetes (chloroindolecarboxamides), inhibit liver Phosphorylase allosterically. These inhibitors bind at the dimer interface, stabilizing the inactive (tense) conformation.
GlcNAc
inhibitor
PLP
PDB 1EM6
Question: Why would an inhibitor of Glycogen Phosphorylase be a suitable treatment for diabetes?
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Glycogenolysis
1. Glycogen phosphorylase Releases glucose only if is 5 units away from branch point Phosphorolytic cleavage of the a(14) glycosidic linkages
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Glycogenolysis
1. Glycogen phosphorylase
Binds a cofactor: PLP (pyridoxal-5-phosphate) Has an: Active state (R = relaxed) Inactive state (T = tense) in which the active site is buried
Glycogen binding site probably allows the enzyme to work many times on the same molecule
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The (16) glucosidase then catalyzes hydrolysis of the (16) linkage, yielding a free glucose.
The major product of glycogen breakdown is glucose-1-phosphate, from Phosphorylase activity.
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Glycogenolysis 3. Phosphoglucomutase G-1-P is catalyzed to G-6-P. Reversible and can lead to glycogen synthesis. In muscle G-6-P coming from glycogen can enter glycolysis and supply energy. Luck of G-6-Phosphatase insures that glucose-6-P can not diffuse out of the cells, therefore it can only be catabolized in glycolysis In liver it can be dephosphorylated and released to the blood. This requires a specific liver enzyme: G-6-P phosphatase.
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Glycogenolysis 3. Phosphoglucomutase In the liver, kidney and intestine )other cells dont have G-6-phosphatase)
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Glycogen
Glucose-1-P
Metabolic Disorders Congenital a) gene is absent b) under expressed c) poorer functioning mutant Acquired a) disease of organ b) disease of endocrine organ
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Glycogen storage diseases are congenital genetic (inherited) enzyme deficiencies that result in excessive glycogen accumulation within cells.
They cause abnormal quantity or quality of glycogen. Result in: Liver: hepatomegaly (enlarged liver) and hypoglycemia (low blood sugar)
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Glycogen storage diseases Type I: Glucose-6-phosphatase deficiency )von Gierkes disease( Increase in cellular G-6-P and accumulation of glycogen in liver and kidney (G-6-P inhibits glycogen phosphorylase). No glycogen breakdown and gluconeogenesis. Inability to increase blood sugar levels in response to glucagon Hepatomegaly, hypoglycemia, small stature (1 in 100,000)
Normal
Treatment to increase blood glucose : * Inhibition of liver glucose uptake (drugs) to increase blood levels * Intragastric feeding overnight to increase blood levels Liver filled * Oral feeding with corn starch that breaks with down slowly to glucose glycogen or *Liver transplantation * or Gene therapy G6Pase knockout mice were treated with a viral vector containing 230 G6Pase - increases survival rate Survival to adult was rare, now typical
Glycogen storage diseases Type V: Muscle phosphorylase deficiency )McArdles disease( Symptoms: Painful muscle cramps Appear at early adulthood Glycogen breakdown system cannot provide enough fuel for glycolysis when there is high demand for ATP (Liver phosphorylase is OK) Prevented by avoiding strenuous exercise Treatment: oral B6 vitamin (PLP) (1 in 100,000)
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Glycogen storage diseases Type VI: Liver phosphorylase deficiency )Hers disease(
Similar symptoms to those with Type I glycogen storage disease Hypoglycemia and hepatomegaly Hypoglycemia because of the inability of glycogen phosphorylase to respond to need for glucose production by the liver
Stunted growth
Good prognosis
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Control of glycogen breakdown: Commonly used terminology: "a" is the form of the enzyme that tends to be active, and independent of allosteric regulators (in the case of Glycogen Phosphorylase, when phosphorylated). "b" is the form of the enzyme that is dependent on local allosteric controls (in the case of Glycogen Phosphorylase when dephosphorylated).
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Allosteric regulation
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Control by phosphorylation
Both hormones are produced in response to low blood glucose. )Epinephrine )adrenaline( is also part of the fight or flight response. (Acute stress response. Reaction of an animal to threats through the nervous system, priming the animal for fighting or fleeing.)
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Control by phosphorylation
Hormone (glucagon or epinephrine) released in response to metabolic state Synthesis of cAMP from ATP by adenylate cyclase P* Activation of protein kinase A (cAMP dependent protein kinase) P*
Glycogen breakdown
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Control by phosphorylation
The cAMP cascade results in phosphorylation of a serine hydroxyl of Glycogen Phosphorylase, which promotes transition to the active (relaxed) state glycogen phosphorelase a. The phosphorylated enzyme is less sensitive to allosteric inhibitors. even if cellular ATP & glucose-6-phosphate are high, Phosphorylase will be active. The glucose-1-phosphate produced from glycogen in liver may be converted to free glucose for release to the blood. With this hormone-activated regulation, the needs of the organism (hormone) take precedence over needs of the cell (cellular ATP).
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Control by phosphorylation
In a resting cell: ATP and G-6-P are usually high enough inhibit phosphorylase b (mostly in T state) Activity of phosphorylase in a resting cell depends on how much is in the phos. a state this depends on how much phosphorylation there is and on the levels of glucose. If glucose is high insulin secretion dephosphorylation it will be in an inactive form (T)
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Control by phosphorylation
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Insulin
Control of glycogen breakdown: Control by phosphorylation Cascade mechanism of epinephrine and glucagon action. The resulting breakdown of glycogen provides glucose, which in the myocyte can supply ATP (via glycolysis) for muscle contraction and in the hepatocyte is released into the blood to counter the low blood glucose. Note the amplification of the signal: from few molecules of hormone to thousands of glucose molecules
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Glycogen synthesis Glycogen synthesis and glycogen breakdown are separate pathways. McArdles disease )Muscle phosphorylase deficiency) showed that glycogen breakdown in muscle is deficient due to lack of glycogen phosphorylase, but glycogen levels were moderately high indicating that phosphorylase was not required for synthesis. Key molecule is UDP-glucose (uridine-diphosphate glucose)
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Glycogen synthesis
UDP-glucose-pyrophosphorylase
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Glycogen synthesis
2Pi
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Glycogen synthesis The reactions are: 1) glucose-1-phosphate + UTP UDP-glucose + PPi 2) PPi + H2O 2 Pi
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Transfers a segment from the end of a glycogen chain to the C6 hydroxyl of a glucose residue of glycogen to yield a branch with an (16) linkage. About 7 glucoses are transferred Branching process increases glycogen solubility and increases a number of non-reducing ends (important for rapid glycogen breakdown)
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Glycogen synthesis
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If both pathways would work at the same time we would have a futile cycle
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Allosteric regulation
Glycogen Synthase is allosterically activated by: glucose-6-phosphate (opposite of the effect on Phosphorylase). glycogen synthesis is activated when glucose-6-phosphate is plentiful. These controls benefit the cell because it is more useful to a cell to store glucose as glycogen when there is enough: input to Glycolysis (glucose-6-phosphate) main product of Glycolysis (ATP)
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Control of glycogen synthesis: Control by phosphorylation Glycogen synthase a active (dephosphorylated) Glycogen synthase b less active (phosphorylated) The cAMP cascade induced in liver by glucagon or epinephrine has the opposite effect on glycogen synthesis = inhibition Glycogen Synthase is directly phosphorylated by cAMP-Dependent Protein Kinase, as well as by Phosphorylase Kinase. Phosphorylation of Glycogen Synthase promotes the "b" (less active) conformation.
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Hormonal effects
Insulin antagonizes the effects of the cAMP cascade induced by glucagon & epinephrine.
Insulin is produced in response to high blood glucose
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Hormonal effects Insulin, produced in response to high blood glucose, triggers a separate signal cascade that leads to activation of Phosphoprotein Phosphatase. This phosphatase catalyzes removal of regulatory phosphate residues from Phosphorylase, Phosphorylase Kinase, & Glycogen Synthase enzymes. Thus insulin antagonizes effects of the cAMP cascade induced by glucagon & epinephrine.
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Hormonal effects Ca++ also regulates glycogen breakdown in muscle. During activation of contraction in skeletal muscle = nerve pulse, the sarcoplasmic reticulum (smooth ER of muscle) releases Ca++. Ca++ released to the cytosol activates actin/myosin interactions and glycogen breakdown (for glycolysis ATP muscle contraction).
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Hormonal effects
Phosphorylase Kinase in muscle includes calmodulin (Calcium-modulating protein) as its subunit. Phosphorylase Kinase is partly activated by binding of Ca++ to this subunit. Calmodulin binds 4 Ca++ ions and the enzyme undergoes a conformational change. Phosphorylation of the enzyme, via a cAMP cascade induced by epinephrine, results in further activation. These regulatory processes ensure release of phosphorylated glucose from glycogen, for entry into glycolysis to provide ATP needed for muscle contraction.
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Hormonal effects Liver: Insulin and glucagon are both made in the pancreas respond to blood glucose levels Muscles and other tissues: Insulin and epinephrine (adrenaline) and nor-epinephrine affect glucose metabolism
Epinephrine induces glycogen breakdown to G-6-P in muscle for glycolysis, or to glucose for export from liver. When there is enough glucose, insulin stimulates glucose uptake by muscle cells and increases glycogen synthesis. Liver cells respond directly to glucose and increase glycogen synthesis. 255
Hormonal effects
Difference in the regulation of carbohydrate metabolism in liver and muscle. In liver, either glucagon (indicating low blood glucose) or epinephrine (signaling the need to fight or flee) has the effect of maximizing the output of glucose into the bloodstream.
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In muscle, epinephrine increases glycogen breakdown and glycolysis, which together provide fuel to produce the ATP needed for muscle contraction.
Glycogen phosphorylase b
Glycogen synthase a
ATP
epinephrine
+
- " phosphoprotein phosphatase
- " phosphoprotein phosphatase " protein kinase
+
" protein kinase
+
Ca+2 AMP
P ATP
Glycogen phosphorylase b
Glycogen synthase a
glucagon
+
- " phosphoprotein phosphatase
- " phosphoprotein phosphatase " phosphorelase b kinase
Glycogen phoshorylase a
Glycogen synthase b
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Glycogen storage diseases Type IV: Branching enzyme deficiency )Andersens disease( Most severe dont live to more than 5 years Type IV glycogen is not increased in liver but the structure is abnormal Very long unbranched glycogen decreases solubility, liver failure
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Gluconeogensis
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Humans consume 160 g of glucose per day 75% of that is in the brain (120 g) Body fluids contain only 20 g of glucose Glycogen reserves yield 180-200 g of glucose The body must be able to make its own glucose when dietary sources of glucose are not available or when the supply of liver glycogen is exhausted Gluconeogenesis In the liver, and less in the kidney
Synthesis of "new glucose" from common metabolites: pyruvate, lactate, glycerol, amino acids and all TCA intermediates
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Gluconeogenesis
The brain and erythrocytes are almost completely dependent on glucose as an energy source.
The glycogen stores in the liver will supply the brain with glucose for half a day of fasting or starvation. When fasting most of the bodys glucose needs must be supplied by gluconeogenesis. Was shown that during a fast: During the first 22 hours 64% of glucose come from gluconeogenesis By 46 hours 100% is from gluconeogenesis Gluconeogenesis is an universal passway and it exists in all organisms
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Gluconeogenesis
Non-carbohydrate precursors: Lactate, pyruvate from glycolysis in skeletal muscle or erythrocytes Amino acids from dietary proteins, or muscle protein after starvation Glycerol from fat breakdown and others Acetyl-coA is not a precursor for gluconeogenesis
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Gluconeogenesis
Synthesis of glucose from pyruvate utilizes many of the same enzymes as glycolysis. Three glycolysis reactions have such a large negative Gs that they are essentially irreversible. These must be bypassed in gluconeogenesis: Hexokinase Phosphofructokinase Pyruvate Kinase.
Therefore gluconeogenesis requires 4 specific enzymes. The other steps use reversible glycolytic enzymes.
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Gluconeogenesis
Glycolysis
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Gluconeogenesis
Glycolysis
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Gluconeogenesis
Need to bypass: pyruvate kinase (pyrPEP) PEP has a higher negative G of phosphate hydrolysis than ATP, so will have to break 2 phosphate bonds to get to PEP. Pyruvate to phosphoenolpyruvate
A. Pyruvate carboxylase
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Gluconeogenesis
Pyruvate to phosphoenolpyruvate
A. Pyruvate carboxylase The enzyme has a biotin prosthetic group. Biotin is an essential nutrient. Biotin functions as a CO2 carrier by forming a carboxyl group at its end. Biotin has a 5-C side chain whose terminal carboxyl is in amide linkage to a lysine amino group in the enzyme. A highly specific enzyme Biotin Protein Ligase catalyzes attachment of biotin to a particular Lys of Pyruvate Carboxylase (and to specific Lys residues in other enzymes that catalyze carboxylation or decarboxylation reactions). The biotin & lysine side chains form a long swinging arm that allows the biotin ring to swing back & forth between 2 active sites (one carboxilates biotin and the other pyruvate)
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Gluconeogenesis Pyruvate to phosphoenolpyruvate A. Pyruvate carboxylase 1) Biotin carboxylation is catalyzed at one active site of Pyruvate Carboxylase: ATP is cleaved to ADP and Pi Pi Reacts with bicarbonate HCO3- to release CO2 CO2 is bound to biotin to form an activated carboxyl
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Gluconeogenesis Pyruvate to phosphoenolpyruvate A. Pyruvate carboxylase 2) At the other active site of Pyruvate Carboxylase, the activated carboxyl is transferred from carboxybiotin to pyruvate to form oxaloacetate.
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Gluconeogenesis Pyruvate to phosphoenolpyruvate A. Pyruvate carboxylase Biotin is an essential nutrient, but deficiency is rare. Avidin, a protein in egg whites with a barrel structure, tightly binds biotin. Excess consumption of raw eggs can cause nutritional deficiency of biotin. The strong avidin-to-biotin affinity is used by biochemists as a specific "glue." To bind proteins together, biotin may be covalently linked to one protein and avidin to the other.
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Gluconeogenesis Pyruvate to phosphoenolpyruvate A. Pyruvate carboxylase Oxaloacetate is: 1) a precursor for gluconeogenesis But is also: 2) An intermediate of citric acid cycle. When the citric acid cycle is inhibited (there is enough ATP), oxaloacetate is sent to gluconeogenesis But oxaloacetate is made by the citric acid cycle in the mitochondria and the next steps are in the cytosol So OAA must be transported out
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NADH levels are low in the cytosol and the cells needs it for gluconeogeneis to proceed. 277
Gluconeogenesis Pyruvate to phosphoenolpyruvate A. Pyruvate carboxylase The first partial reaction of pyruvate carboxylase, the formation of carboxybiotin, depends on the presence of acetyl-CoA. Biotin is not carboxylated unless acetylCoA is bound to the enzyme. Pyruvate Carboxylase is allosterically activated by acetyl-CoA. Acetyl-CoA has no effect on the second partial reaction. If acetyl-CoA is high, it signifies the need for more oxaloacetate Then, on the cell level: 1) If the energy charge is high, it means that there are enough substrates for the citric acid cycle, and oxaloacetate is converted into glucose (gluconeogenesis). 2) If the energy charge is low, oxaloacetate replenishes the citric acid cycle.
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Then, on the cell level: 1) If the energy charge is high, it means that there are enough substrates for the citric acid cycle, and oxaloacetate is converted into glucose (gluconeogenesis). 2) If the energy charge is low, oxaloacetate replenishes the citric acid cycle.
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Gluconeogenesis Pyruvate to phosphoenolpyruvate B. Phosphoenolpyruvate carboxykinase (PEPCK) The reaction is thought to proceed in two steps: Oxaloacetate is first decarboxylated to yield a pyruvate enolate anion intermediate. This is phosphorylated by phosphate transfer from GTP. A metal ion such as Mn++ is required, in addition to the Mg++ associated with the nucleotide substrate.
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A. Pyruvate carboxylase
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Gluconeogenesis Specific enzymes for bypass reactions: Hydrolytic reactions, no ATP Release of Pi in an exergonic process 1) Fructose-1,6-bisphosphate to Fructose-6-phosphate by Fructose-1,6-bisphosphatase 2) Glucose-6-phosphate to Glucose by Glucose-6-phosphatase (used in glycogen breakdown)
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Gluconeogenesis Gluconeogenesis occurs only in liver and kidney because of the presence of Glucose-6-phosphatase. Only these tissues supply glucose to other tissues. Muscle and brain do not do gluconeogenesis G-6-phosphatase is found in the ER (active site is exposed to cytosol) G-6-P is hydrolyzed as it passes into the ER and glucose enters the ER ER vesicles filled with glucose diffuse to the plasma membrane, fuse with it and open, releasing glucose into the bloodstream.
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Gluconeogenesis Energy expenses: Anabolic pathways cost energy Glycolysis & Gluconeogenesis are both spontaneous (due to their irreversible steps). Glycolysis: glucose + 2 NAD+ + 2 ADP + 2 Pi 2 pyruvate + 2 NADH + 2 ATP Gluconeogenesis: 2 pyruvate + 2 NADH + 4 ATP + 2 GTP glucose + 2 NAD+ + 4 ADP + 2 GDP + 6 Pi
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Gluconeogenesis
If both pathways were simultaneously active in a cell, it would constitute a "futile cycle" that would waste energy.
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Regulation of gluconeogenesis Glycolysis and gluconeogenesis are reciprocally controlled Regulation occurs on the strongly exerogenic processes in each pathway the non-reversible enzymes * When glycolysis is turned on gluconeogenesis should be turned off
High energy status: glycolysis off Pyruvate and other substrates are used for synthesis and storage of glucose
Low energy status: glycolysis on, to rapidly degrade glucose and supply energy
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(Gs)
When cellular ATP is high (low AMP), glucose is not degraded to make ATP. Under such conditions the cell prefers to store glucose as glycogen. Phosphofructokinase (Glycolysis) is inhibited by ATP and stimulated by AMP. Fructose-1,6-bisphosphatase (Gluconeogenesis) is inhibited by AMP. ATP regulates pyruvate kinase in liver cells: If ATP is high it inhibits glycolysis and induces gluconeogenesis that occurs in liver.
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Regulation of gluconeogenesis
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Regulation of gluconeogenesis
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Regulation of gluconeogenesis
Glucose-6-phosphatase is under substrate-level control, not allosteric control The Km of the enzyme glucose-6 phophatase is higher than intracellular levels of G6P, and will then work when the levels are high.
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Fructose-2,6-bisphosphate is not a glycolytic intermediate but controls both pathways. F-2,6-bisphosphate: 1) a potent activator of PFK (PFK1) from glycolysis 2) an inhibitor of Fructose 1,6 BPase from gluconeogenesis F-2,6-bisphosphate is synthesized and degraded by the same molecule with two enzymatic activities: PFK2 and fructose bisphosphatase-2
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Regulation of gluconeogenesis
Global control in liver cells includes reciprocal effects of a cAMP cascade, induced by glucagon when blood glucose is low. Phosphorylation of enzymes by cAMP-Dependent Protein Kinase results in inhibition of Glycolysis & stimulation of Gluconeogenesis, making glucose available for release to the blood.
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Pyruvate Kinase (Glycolysis) is also inhibited when phosphorylated via cAMP-Dependent Protein Kinase
Regulation of gluconeogenesis
Phosphofructokinase (PFK-1) is normally activated by fructose-2,6-bisP, so a cAMP-induced decrease in [fructose-2,6-bisP] slows this Glycolysis enzyme.
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Glycogen
X
Glucose-1-P
Glycolysis Pathway
Summary of effects of glucagon-cAMP cascade in liver: Gluconeogenesis is stimulated. Glycolysis is inhibited. Glycogen breakdown is stimulated. Glycogen synthesis is inhibited. Free glucose is formed for release to the blood.
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Regulation of gluconeogenesis
The Cori Cycle connects between muscle and liver It operates during exercise, when aerobic metabolism in muscle cannot keep up with energy needs.
For a brief burst of ATP utilization (2 to 7 sec), muscle cells utilize ~P stored as phosphocreatine. For more extended exercise, ATP is mainly provided by Glycolysis.
In order to avoid a futile cycle in one cell glycolysis occurs in muscle and gluconeogeneis in the liver
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Regulation of gluconeogenesis
Vigorous exercise can lead to a buildup of lactate and NADH, due to oxygen shortage and the need for more glycolysis NADH can be reoxidized during the reduction of pyruvate to lactate Lactate is then returned to the liver through the bloodstream, where it can be reoxidized to pyruvate by liver Lactate Dehydrogenase
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The Cori cycle costs 6 ~P in liver for every 2 ~P made available in muscle. The net cost is 4 ~P. Although costly in ~P bonds, the Cori Cycle allows the organism to accommodate to large fluctuations in energy needs of skeletal muscle between rest and exercise.
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Gluconeogenesis Pyruvate to phosphoenolpyruvate There is another bypass from pyruvate to PEP when lactate is the glucogenic precursor. When lactate dominates (after exercise) the conversion of lactate to pyruvate in the cytosol of hepatocytes yields NADH, and the export of reducing equivalents (as malate) from mitochondria is therefore unnecessary.
The oxaloacetate is then converted directly to PEP by a mitochondrial isozyme of PEP carboxykinase, and the PEP is transported out of the mitochondrion to continue on the gluconeogenic path.
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Outline of pathways for glucose synthesis from the major gluconeogenic precursors.
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Gluconeogenesis Major gluconeogenic precursors glycerol TG breakdown yields fatty acids and glycerol. Glycerol can be used in gluconeogenesis converts to glycerol-3-P first and then to DHAP.
Fatty acids oxidation yields acetyl-coA not a gluconeogenetic precursor (unless there is glyoxylate cycle)
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Animations
http://www.wiley.com/legacy/college/boyer/0470003790/animations/gluconeoge nesis/gluconeogenesis.swf
http://www.wiley.com/legacy/college/boyer/0470003790/animations/cori_cycle/cori_c ycle.htm
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Phenylketonuria examination
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Vitamin D
Vitamin E
Vitamin K
K1
K2
Vitamin C
GOUT
Uric acid
Excretion of bilirubin
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