ALTERNATIVE PATHWAY FOR CARBOHYDRATE METABOLISM (for clearer images, pls. refer to the ppt) Question: How many ATPs do you need in the conversion of FRUCTOSE-6-PHOSPHATE to PYRUVATE? Answer: 1 ATP only since we have already bypassed the major regulatory pathway of glycolysis which is PHOSPHOFRUCTOKINASE 1 It is not recommended to carbo load on fructose since it cannot be controlled (Phosphofructokinase regulates the amount that the body requires) -This pathway is used as an alternative when there is a high demand for NADPH which is required in FATTY ACID SYNTHESIS TISSUE DISTRIBUTION -Demand for NADPH • Biosynthetic pathways -fatty acid synthesis (liver, adipose, mammary) -cholesterol synthesis (liver) -steroid hormone synthesis (adrenal, ovaries, testes) • Detoxification (Cytochrome P450 system)  liver • Reduced glutathione as an antioxidant in RBC to maintain the shape of RBC -preventing hemolysis • Generation of superoxide (neutrophils) **All these conditions will cause an INCREASE in NADPH demand which can lead to the hexose monophosphate shunt FUNCTIONS -Source of: 1. NADPH + H+  for REDUCTIVE BIOSYNTHETIC PROCESSES (synthesis of cholesterol, fatty acids, steroid hormones) -more important function 2. Pentoses  for nucleotides, nucleic acids and coenzymes CHARACTERISTICS -neither consumes nor produce ATP -that is why it is not the major pathway -branches off glycolysis at G6P (common to both glycolysis and HMP shunt) -most important molecule in this pathway is GLUCOSE-6PHOSPHATE -glucose should first be converted to glucose-6phosphate before entering the hexose monophosphate shunt -GLUCOKINASE in the liver and HEXOKINASE in the muscles  enzymes needed for the conversion of glucose to glucose-6-phosphate -common enzyme in glycolysis and HMP shunt -re-entry is at FRUCTOSE-6-PHOSPHATE -PHOSOHEXOSE ISOMERASE enzyme required for the conversion of glucose-6-phosphate to fructose-6-phosphate

SUBJECT: BIOCHEMISTRY TOPIC: CARBOHYDRATE METABOLISM 3 LECTURER: DR. ESPERANZA UY DATE: DECEMBER, 2010

Question: How many ATPs do you need in the conversion of SUCROSE to PYRUVATE? (It will come out in the exam.) Answer: 3 ATPs. Sucrose is formed by glucose and fructose. From glucose to pyruvate, 2 ATPs are needed. For fructose to pyruvate, 1 ATP is required. In conclusion, 3 ATPs are required since 2 ATPs are needed for the breakdown of glucose and 1 ATP is needed for the breakdown of fructose ALTERNATIVE PATHWAY OF METABOLISM -Pentose Phosphate Pathway -Uronic Acid Pathway -Fructose Metabolism -Metabolism of Galactose -Sorbitol Pathway -Ethanol Metabolism -Hexosamine Formation  HEXOSE MONOPHOSPHATE SHUNT -aka PENTOSE PHOSPHATE PATHWAY -an ALTERNATE ROUTE for glucose oxidation -not the main main pathway -major pathway is GLYCOLYSIS -found in the CYTOSOL -active in: liver, adipose tissue, lactating mammary gland, adrenal cortex and RBC -common among these organs: all are secretory organs, except for RBC -5 carbons are produced in this pathway -the 1 carbon will be produced as CO2 -the only pathway that produces CO2 in cytoplasm -carbon dioxide is normally produced in the mitochondria but in this pathway, the carbon dioxide is in the cytoplasm -NADPH is important in this pathway (PPP is the major source of NADPH in cells) BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3

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CHARACTERISTIC DIFFERENCE WITH GLYCOLYSIS -oxidation utilizes NADP rather than NAD -carbon dioxide is a characteristic product of HMP shunt -only source of carbon dioxide in the cytoplasm -increase demand for NADP happens where there is an increase in demand for fatty acid synthesis -NO ATP is generated PENTOSE PHOSPHATE PATHWAY HAS 2 PHASES A. Glucose-6-Phosphate is oxidized and decarboxylated to a pentose sugar (generates 2 moles of NADPH per G6P oxidized) -irreversible

NON-OXIDATIVE PHASE -starts in the ribulose-5-phosphate molecule -reversible (can go anywhere) -rearrangement of sugars to enter glycolytic pathway -provides a pathway for recycling excess pentoses IMPORTANT ENZYMES: (refer to the diagram below as well) 1) 3-epimerase -to convert ribulose-5-phosphate to xylulose-5-phosphate **epimerase  translocation in only carbon 2) Ketoisomerase -to convert ribulose-5-phosphate to ribose-5-phosphate 3) Transketolase -will transfer 2 carbon atoms from xylulose-5-phosphate to ribose-5-phosphate to form glyceraldehyde-3-phosphate and sedoheptulose-7-phosphate -important coenzyme: THIAMIN (Vit. B1) -important cofactor: MAGNESIUM (Mg+) -source: unpolished rice ** blocking transketolase activity may lead to pentosuria (a pentose sugar, usually xylulose, is found in urine ** thiamine deficiency affects sedoheptulose 7 phosphate since it affects transketolase activity too 4) Transaldolase -will transfer 3 carbon atoms -sedoheptulose-7-phosphate is a very unstable compound that is why it will be acted upon by TRANSALDOLASE to form erythrose-4-phosphate and fructose-6-phosphate which will then be converted to glucose-6-phosphate through the enzyme PHOSPHOHEXOSE ISOMERASE **Another TRANSKETOLASE (similar with the transketolase mentioned above) will act on xylulose-5-phosphate to be converted to FRUCTOSE-6-PHOSPHATE and GLYCERALDEHYDE-3-PHOSPHATE 5) Phosphotriose isomerise -converts glyceraldehydes-3-phosphate to fructose-1,6bisphosphate 6) Fructose-1,6-bisphosphatase -also found in GLUCONEOGENESIS because it uses the fructose-1,6-bisphosphate as a substrate -converts fructose-1,6-bisphosphate to fructose-6phosphate **all the other enzymes are also found in the glycolysis pathway except the FRUCTOSE-1,6-BIPHOSPHATASE which is found in the gluconeogenesis pathway

**GLUCOSE-6-PHOSPHATE  substrate used in HMP shunt -fructose-6-phosphate and glycogen can also be used as a substrate but they should first be converted to glucose-6-phosphate B. Interconversions of pentose phosphates lead to intermediates of glycolysis – reversible **FRUCTOSE-6-PHOSPHATE and GLYCERALDEHYDE  important intermediates of glycolysis OXIDATIVE PHASE -irreversible -generates NADPH  2 moles per glucose oxidized -3 reactions  starts with glucose-6-phosphate and results in ribulose-5-phosphate **3 glucose-6-phosphate molecules are required **NADPH producing reactions/ important enzymes needed in the oxidative phase: • Glucose-6-phosphate dehydrogenase -coenzyme: NADP -water is also needed to be reduced to NADPH + H -product: 6-PHOSPHOGLUCONATE • 6-phosphogluconate dehydrogenase -NADP is required in this step -decarboxylates 6-phosphogluconate which RELEASES CARBON DIOXIDE from the cytoplasm -product: ribulose-5-phosphate Question: Why do we need 3 glucose-6-phophate molecules? Answer: To produce 2 fructose-6-phosphate molecules and 1 glyceraldehyde molecule through the interconversion of the pentoses

BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3

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GLUCOSE-6-PHOSPHATE DEFICIENCY -problem in glucose-6-phosphate dehydrogenase enzyme -not evident in most cases -common in the Philippines (usually affects males, while women are carriers) -induced by drugs:    Antimalarials sulfa antibiotics antipyretics (aspirins,etc.)

-acute haemolytic anemia is due to decreased NADPH production -HMP is the major pathway of NADPH production in the red blood cell (G6PD deficient RBCs = more prone to hemolysis) -NADPH is responsible for maintaining glutathione in its reduced state -reduced glutathione is necessary for the integrity of the erythrocyte membrane, thus rendering enzyme-deficient red cells more susceptible to hemolysis by a wide range of compounds **1/2 glucose-6-phosphate glyceraldehyde **3 glucose-6-phosphate molecules are required, 6 molecules of NADPH + H+ are produced and 3 CO2 from the cytoplasm are released to have the product, xylulose-5phosphate  irreversible **the reversible process will ultimately produce 2 molecules of glucose-6-phosphate and 1 molecule of glyceraldehydes. All of the three molecules will enter the glycolytic pathway -entry point GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE **defiency in thiamine (VIt.B1)  problem in transketolase enzyme. This can cause hemolytic anemia and jaundice OVERALL EQUATION: Glucose -6- PO4 + 2 NADP+ + H2O ↔ ribose -5- PO4 + 2NADPH + 2 H+ + CO2 REGULATION 1.Glucose-6-phosphate dehydrogenase -first step; where regulation acts most -rate limiting step -most important regulator -abundant in the RBC 2.Allosteric regulation -feedback inhibited by NADPH -increase amounts of NADPH will cease the HMP shunt -increased NADPH also entails and increased amount of H ion which decreases intracellular pH (becomes more acidic); can shrink and wrinkle the cell 3.Inducible enzyme -induced by insulin -also found in the glycolytic and glycogenesis pathway -increased amounts of insulin can induce the HMP shunt (since much glucose needs to be distributed) Question: In the formation of lactate from glucose-6phosphate, how many ATPs are needed? Answer: 2 ATPs BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3 Page 3 -exposure to anti-malarial drugs (Primaquine) results in increased cellular production of superoxide and hydrogen peroxide (Primaquine sensitivity) -other chemicals known to increase oxidant stress -sulfonamides (antibiotic) -aspirin and NSAIDs -quinidine and quinine -naphthalene (mothballs) -fava beans (vicine and isouramil) ROLE OF NADPH IN THE RBC 1.Production of superoxide 2.Hb-Fe2+-O2  Hb-Fe3+ + O23.O2- + 2H2O  2H2O2 4.Both O2- and H2O2 can produce reactive free radical species, damage cell membranes and cause hemolysis DETOXIFICATION OF SUPEROXIDE ANION AND HYDROGEN PEROXIDE (H202) -Antioxidant enzymes -superoxide dismutase -glutathione peroxidise -if the glutathione is in the reduced state, hydrogen peroxide will be converted to water -glutathione reductase

 THE URONIC ACID PATHWAY -also encountered in HEME METABOLISM (for more details, see Bilirubin Conjugation) -catalyzes the conversion of glucose to glucuronic acid, ascorbic acid and pentoses **GLUCORONIC ACID  to conjugate bilirubin so that it can be more soluble in water -also an alternative oxidative pathway for glucose -does not lead to ATP formation G6P  G1P  UDP-Glucose  UDP- Glucoronic Acid Glucoronic Acid  L-Gulonic Acid (a direct Vit. C precursor as well)  3-Keto-L-Gulonic Acid (+CO2) L-Xylulose Xylitol (usual candy component)  D-Xylulose  D-Xylulose-5-P **conversion of glucose-6-phosphate to glucose-1phosphate was also encountered in glycogenolysis **PENTOSURIA  blocks the L-xylulose  xylitol pathway -causes the presence of xylulose in the urine **xylulose-5-phosphate will then enter the HMP shunt **The L-gulonate has 2 pathways: 1 for the formation of xylulose-5-phosphate and the other for the formation of ascorbic acid. -However, the formation of ascorbic acid is not possible in higher forms of life since the L-gulonolactone 2ketogulonolactone pathway is BLOCKED. So in humans, the only possible route is for it to be converted to xylulose-5phosphate which will now enter the HMP shunt -this means that Vitamin C cannot be produced by the body and supplements should be taken in to supply the body with Vit.C -high amounts of Vit.C in the body can lead to stones

GLUCORONIDES FORMATION

GLUCURONIC ACID: PHYSIOLOGICAL SIGNIFICANCE OF GLUCURONIDE FORMATION -glucuronide formation is important during detoxification, steroid excretion, and bilirubin metabolism -the reaction is catalyzed by UDP-glucuronyl transferase, which may take several days to 2 weeks after birth to become fully active in humans -babies are always exposed to the sun to hasten the process of bilirubin solubility  FRUCTOSE METABOLISM ESSENTIAL FRUCTOSURIA -fructokinase is deficient in essential fructosuria, a benign asymptomatic metabolic anomaly; autosomal recessive -following intake of fructose, blood and urinary fructose levels of affected individuals are unusually high; however, 90% of their fructose intake is eventually metabolized

**Fructose can also be a source of glucose through the enzyme PHOSPHOFRUCTOISOMERASE **hexokinase is still needed to form fructose-6-phosphate **Aldolase A used in glycolysis **Aldolase B  used in fructose metabolism **Fructokinase  deficient in essential fructosuria -no formation of fructose-1-phosphate and instead, fructose from diet will be converted in the glycolysis pathway (not that significant) BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3 Page 4

HEREDITARY FRUCTOSE INTOLERANCE -deficient aldolase B enzyme; autosomal recessive -characterized by severe hypoglycaemia after ingestion of fructose, and prolonged ingestion by affected young children may lead to death -fructose-1-phosphate aldolase is deficient and fructose 1phosphate accumulates intracellularly -causes cataract formation -will eventually go to the glycolytic pathway

GALACTOSEMIA -inability to transform galactose into glucose -individuals with this defect are unable to metabolize the galactose derived from lactose (milk sugar) to glucose metabolites -can lead to cataract formation, growth failure, mental retardation or death from liver failure -may be due to deficiency of: -GALACTOKINASE  leads to cataract formation -GALACTOSE-1-PHOSPHATE URIDYLTRANSFERASE leads to severe disease (liver disease) **ALDOSE REDUCTASE which is abundant in the lens, liver, mammary glands and kidneys -through the NADPH from HMP shunt converts galactose to GALATITOL which can cause cataracts -not a significant enzyme, but only becomes important if galactose levels are very high and a deficiency in galactokinase - Aldose Reductase Inhibitors = known side effects are severe cardiovascular and GIT problems

 METABOLISM OF GALACTOSE -from LACTOSE -lactose  glucose + galactose -abundant in human milk -UDP-galactose is also formed from free galactose derived from hydrolysis of lactose in the intestinal tract -galactose is phosphorylated by galactokinase and ATP to yield galactose-1-phosphate -galactose-1-phosphate uridyltransferase froms UDPgalactose from galactose 1-phosphate displacing glucose 1-phosphate from UDP-glucose

> Classic Galactosemia – more severe than galactosemia deficiency; patient may die at womb  SORBITOL / POLYOL PATHWAY -not found in the liver -active in those tissues that are not insulin-sensitive -lens -peripheral nerves -renal glomeruli -maybe involved in the pathogenesis of diabetic cataracts and diabetic peripheral neuropathy **1 ATP is required to form GALACTOSE-1-PHOSPHATE **galactose-1-phosphate uridyl transferase came from UDP-glucose from GLYCOGENOLYSIS PATHWAY -blocked in galactosemia BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3 -can have effects on the liver, lens and seminal vesicle (can lead to infertility) -found in the cytosol **NADPH is needed Page 5

 METABOLISM OF HEXOSAMINE OR AMINO SUGARS -another mechanism in diabetic patients -JUST REMEMBER: from glycogen, glycosaminoglycans, sialic acid, gangliosides and glycoproteins in the brain can be formed COMPLEX POLYSACCHARIDES -Glycoproteins **sorbitol dehydrogenase is not a very important enzyme but only becomes essential if there is an increased amount of glucose in the body **sorbitol pathway is NOT FOUND IN THE LIVER **active in tissues that are not insulin sensitive **”UNIFYING PATHWAY OF DIABETICS” **sorbitol attracts a lot of water Night: Glycogenolysis: Glucose Fasting  inactive sorbitol pathway = clear eyesight Day: Gluconeolysis: Glucose Usage  active sorbitol pathway = unclear eyesight (due to trapping in lens) -Proteoglycans GLYCOPROTEINS -conjugated proteins containing one or more saccharides LACKING A SERIAL REPEAT UNIT and are bound covalently to a protein -important in the biological functions of the membrane -constitutes of the mucus -IMPORTANCE: lubrication and protection of the tissues lining the respiratory, GIT, and female reproductive system -examples: hormones, plasma proteins PROTEOGLYCANS -contains 95% or more of carbohydrates -carbohydrate chains are called GLYCOSAMINOGLYCANS or MUCCOPOLYSACCHARIDES -present in connective tissues -6 classes: -chondroitin sulfate -dermatan sulfate -keratin sulfate -heparan sulfate -heparin -hyaluronate GLYCOSAMINOGLYCANS (GAGs) Common Features 1. GAGs are made up of dissacharide repeating units, hexosamine and a uronic acid; highly (-) charged  ETHANOL METABOLISM Oxidation to acetate in the liver -ethanol is oxidized in the liver by CYTOSOLIC ALCOHOL DEHYDROGENASE to acetaldehyde -the acetaldehyde is further oxidized to acetate by MITOCHONDRIAL ALDEHYDE DEHYDROGENASE -much of the acetate produced from ethanol leaves the liver and is converted by acetyl coA, which can be used to provide energy via the TCA cycle -alcohol will be used for ATP production -acetyl coA may also be formed in the liver and used as a precursor for lipid biosynthesis BIOCHEMISTRY: CARBOHYDRATE METABOLISM 3 2. they contain sulfate groups linked by ester bonds to certain monosaccharides orby amide bonds to the amino group of glucosamine 3. only hyaluronate is not sulphated and is not attached to protein 4. the carboxyls of uronic acids and the sulfate groups contribute to the highly charged nature of GAGs 5. predominantly components of the extracellular matrices and cell surfaces, and they participate in cell adhesion and signalling **GAGs are destroyed in diabetic nephropathy -------------------------------------END OF TRANX-------------------------------------

  GOODLUCK ON YOUR EVALUATIONS, BATCH 2014!  

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