Medical Biochemistry

Second Year Program I Semester (Revised in December, 2014)

1. Contractile proteins of skeletal muscle. Tropomyosin and troponin

- Skeletal muscle basics
- STRIATIONS
 Longitudinal striations = myofibrils in the muscle fiber
 Transverse striations = due to varied contractile protein composition in the myofibril
- Sarcolemma
- Sarcoplasm  myofibrils running through, droplets of triacylglycerol + glycogen, myoglobin, myogen, myoalbumin,
multinucleated, mitochondria
- Sarcoplasmic reticulum = closed tubular membrane system that extends throughout the sarcoplasm and surrounds
the bundles of contractile proteins within each myofibril
o Longitudinal sections run the length of each sarcomere
o Terminal cisternae that are associated with T-tubules to form a triad
- T tubules = invaginations of the cell membrane, associated with the terminal cisternae, considered to be an
extracellular space, @ Z band or closer to the A band and I band junction

- SARCOMERE
 Alternating light + dark bands when viewing a
longitudinal cross-section
 Dark bands = A band (anisotropic, birefringent),
actin + myosin
 Light bands = I band (isotropic, doesn’t alter
polarized light), actin only
 Other elements:
o Z disc/band = @ center of I band, one Z disc
to another is 1 sarcomere, contains
vimentin and desmin
o H band/zone = @ center of A band, only tails of myosin
o M line = bisects H zone, contains myomesin and creatinine kinase
 Sarcomere is composed of thin + thick filaments (actin and myosin)  6 thin filaments around 1 thick filament, 3
thick filaments around 1 thin filament

- CONTRACTILE PROTEINS
 Myosin
o Makes up thick filament and about 400 myosin rods are bound to together in 1 filament
o Most abundant protein in skeletal muscle (60-70%)
o 2 identical heavy chains and 2 pairs of light chains (i.e. 4 light chains)
 Heavy chains  twisted to make the myosin tail
 Light chains  forms 2 myosin heads
o Myosin has a double-headed globular region is joined to a double stranded  helical rod
o Globular portions of the myosin have ATPase activity + can bind actin
o 2 of the light chains are identical (one on each globular head and may be removed without loss of ATPase
activity
o Other 2 aren’t identical and loss = loss of ATPase activity and ability to bind actin
o The region between the tail and globular head is thought to be flexible
o Midpoint @ which the myosin fibers meet tail to tail is at the M line located in the H zone
 Actin
o Makes up thin filament
o 20-25% of total muscle protein
o Globular form = G-actin, consists of a
single peptide chain, contains myosin-
binding site
o F-actin = double stranded helix of G-
actin monomers
 Tropomyosin
o Rod shaped
o Associated with actin
o Consists of 2 similar -helical peptide
chains coiled around each other
o Attached noncovalently to actin
o Coils around actin and covers the
myosin-binding sites
  Troponin
o Spherical molecule
o Associated with actin
o 3 subunits:
1. TnC = Ca binding protein, 2 high and 2 low affinity Ca binding sites (ONLY subunit that binds Ca)
2. TnI = inhibits actin and myosin interaction and inhibits ATPase activity
3. TnT = tropomyosin binding subunit, mediates binding of both TnC and TnI to actin-tropomyosin
complex + binding of TnI to TnC
o When Ca binds to the TnC  conformational change recognizes by both tropomyosin and actin that
leads eventually to contraction
 -actinin = anchors thin filaments to the Z disc/band
 -actinin = associated with F-actin and is a length-determining factor for thin filament assembly
 -component/creatinine kinase = catalyzes P group transfer from phosphocreatine to ADP, forms ATP for
contraction
 C-protein = involved in assembly of myosin into thick filament
 M-protein = detected in M line
 Titin = anchors myosin to Z disc
 Nebulin = binds to each thin filament and helps to anchor it to -actinin

2. Muscle contraction: Role of the sarcoplasmic reticulum in muscle contraction and relaxation

- MUSCLE CONTRACTION
 Muscle contraction involves shortening of the sarcomere
 Occurs when actin filaments slide past the myosin
filaments towards the M line  myosin heads pull the
thin filaments towards the M line/into the H zone
 Sliding is facilitated by: ATPase activity of globular head of
myosin, its actin binding capactiy and the way in which
myosin aggregates to form the thick filament
 There is a directionality to contraction evident by the
opposite orientations of the globular heads of myosin in
regards to the M line which explains why sliding occurs
towards the M line
 Actin binding to myosin head  cross-linkage/cross-
bridge, exerts a force on one side of the A band that is opposite to that on the other side of the A band = thick
filament is bipolar
 Thin filaments also exhibit polarity
 Myosin is degraded by proteolytic enzymes (e.g. chymotrypsin) into:
o Heavy meromyosin (HMM) = globular heads connected to
short portions of the -helical tails (“neck” of myosin)
o Light meromyosin (LMM) = remaining portion of the -
helical tails
 Particles of HMM combine with actin filaments with the  helical
portions of the HMM orientated in the same angular direction
 Particles of HMM combine with actin filaments that are attached to
the Z line with the -helical portions orientated in opposite angular
directions
 Therefore, since the thin filaments on one side of the Z line have the same orientation and those on the other
side of the Z line have reverse polarity/or are in the opposite direction, the force generated by actin + myosin
binding is in the direction of the M line
 Process of muscle contraction:
o Electrical impulse from motor nerve arrives @ NMJ
o Stimulates release of Ach gets released across the synaptic cleft
o Ach binds to nicotinic acetylcholine receptors allowing influx of Na ions
o The impulse spreads over the entire sarcolemma, which then becomes depolarized
o Potential difference = 60 mV between inside and outside of the resting muscle cell
o As the impulse spreads, the potential disappears as Na enters the sarcoplasm with K loss to outside the
cell
o T tubule becomes depolarized as well (invagination of the sarcolemma) and transmits the impulse to all
of the myofibrils within the muscle fiber
o Since the T tubules are associated with the terminal cisternae of the SR (arranged in triad), the impulse is
also transmitted to the T tubules
o Ca2+ is discharged into the sarcoplasm from the SR (efflux of Ca2+ involves phosphorylation of ATPase and
opening of Ca2+ channels)
o Ca concentration in the sarcoplasm increases 10x
o Ca2+ will then bind to TnC subunit of troponin  conformational change recognized by tropomyosin
 o Tropomyosin moves to expose the myosin binding sites on the actin
o In the resting state, myosin binds ATP which alters the configuration of the myosin heads so that it
releases actin
o The subsequent hydrolysis of ATP to ADP + Pi via myosin’s ATPase activity makes myosin assume a
position ready for contraction and also increases the affinity of actin towards myosin
o So then when the tropomyosin block gets removed, actin will combine with the ADP, Pi-myosin complex
to form actomyosin with the release of ADP and Pi
o Myosin moves along actin filament/slides = contraction towards the M line/H zone
o Other notes:
 The splitting of ATP to ADP powers the muscle contraction
 Actin has a high affinity towards myosin and for the myosin, ADP, Pi-complex but a low affinity
for the myosin-ATP complex
 Hence actin alternately binds to myosin and will be released from myosin as ATP is hydrolyzed
 The force generating step involves a conformation change in the flexible region between the
globular head and the myosin tails
o Relaxation of the muscle occurs with cessation of the nervous impulse and the return of the sarcolemma
to its original polarized state
o Ca2+ is transported back into the SR by the ATP-dependent Ca2+ pump
 ROLE OF SR IN MUSCLE CONTRACTION + RELAXATION
o The sarcoplasmic reticulum functions as a Ca2+ storage reserve
o SR from skeletal muscle is readily isolated and will reseal into closed vesicles that can be isolated by
centrifugation
o The vesicles retain the ability to pump Ca2+ coupled to ATP hydrolysis by the Ca2+-dependent ATPase
o 4 proteins are found in the SR:
1. Ca2+, Mg2+-dependent ATPase has 2 high affinity Ca binding sites, forms phosphoprotein
intermediates when hydrolyzing ATP and its decomposition is activated by Mg 2+
2. LMW proteolipid  rich in arginine and glutamic acid bonded to 2 fatty acids, involved in
mediating Ca transport across the lipid bilayer
3. 2 Ca binding proteins  no enzymatic activity
 An acidic protein called calsequestrin has one half of the amino acid residues as
glutamic and aspartic acid
 Another Ca binding protein is less acidic and binds about one half as much Ca 2+ as
calsequestrin
o Ca2+ transport system
 ATPase + LMW proteolipid function as intrinsic membrane proteins
 ATPase has both polar and non-polar domains  non polar portion is in the lipid bilayer and
contains a Ca2+ ionophore site, polar portion is on the surface of the membrane and is the site for
ATP hydrolysis
 Interaction between the site of ATP hydrolysis and the non-polar portion within the membrane
controls Ca transport across the membrane
 The 2 Ca2+-binding proteins are extrinsic proteins bound to the membrane inside the SR lumen
and stores the Ca2+ in the SR

3. Energy source for muscle contraction

- Derived from ATP hydrolysis
- Concentration of ATP in resting muscle can supply sufficient energy for contraction for only a fraction of a second
- The energy demands of intense contraction will exceed the capacity of muscle to generate sufficient ATP
- Therefore, muscle contains a reserve store of “high energy phosphate” in the form of phosphocreatine
- Resting muscle contains 6x more phosphocreatine than ATP
- When usage of ATP exceeds production, which is what happens during intense
contraction  phosphocreatine stores are used to replenish ATP, considered a backup
system
- So when ATP is needed, ADP is phosphorylated by phosphocreatine catalyzed by
creatine kinase (phosphocreatine + ADP  creatine + ATP)
- When completing a contraction OR when the generation of ATP exceeds its usage,
phosphocreatine kinase catalyzes the phosphorylation of creatine by ATP
- There’s also a second backup system for generating additional ATP  myokinase (2 ADP  ATP + AMP)

4. Metabolic fuel of skeletal muscle after a meal

- Metabolic fuel after a meal = glucose
- After a meal glucose and insulin levels in the blood are high, free fatty acid
levels are low
- Glucose is metabolized mainly via glycolysis but a small proportion (2%) is
metabolized by the HMP shunt
- Use of glucose in different situations:
- Resting muscle:
 Glycolysis and TCA cycle operate @ 10 and 5-10% maximal capacity
  Most of the products of glycolysis are completely oxidized in noncontracting muscle with only small amounts
of lactate entering the blood
- During anoxia:
 Rates of glucose uptake + lactate production are stimulated in anoxia
 Since in anoxia, both ATP and phosphocreatine levels are low and AMP is high, the glycolytic pathway gets
activated because these factors activate PFK, pyruvate kinase and GA3P DH
 The enzyme PFK is also activated by other compounds found in increased levels in anoxic muscle
 During anoxia glycogen stores can also be used but muscle can’t use fatty acids or ketone bodies for energy in
anoxia
- Contracting muscle:
 Consumption of glucose and oxygen increases 20x
 Stimulation of glycolysis  same mechanism as in anoxia
 Stimulate of TCA cycle  increased activity of isocitrate DH
- Prolonged heavy contraction:
 Rate of pyruvate production by glycolysis exceeds its rate of metabolism by the TCA cycle and large amounts of
lactate are formed and are transported to the liver
 Fate of lactate remaining muscle after contraction has not been known for a while BUT now they’ve
determined that:
o Fast-twitch red and white types of skeletal muscle  synthesize glycogen from lactate
o Slow-twitch red type of skeletal muscle  limited capacity of glycogen synthesis due to lack of F-1,6-BP

5. Metabolic fuel of skeletal muscle in fasting state and in tetanic contraction

- FASTING
 Metabolic fuel during early states of fasting = free fatty acids + BCAA
o The FFA are mobilized from adipose tissue
o Fatty acids supply at least 50—60% of the energy needs of skeletal
muscle both at rest and in contraction
o Presence of free fatty acids suppresses the uptake and oxidation of
glucose
o Use of FA by muscle doesn’t require insulin
o The BCAAs (leucine, isoleucine and valine) are degraded in fasting
for energy in extrahepatic tissues (notably skeletal and cardiac
muscle)
o Both FA and epinephrine stimulate oxidation of the BCAAs by skeletal muscle
o Action of epinephrine on skeletal muscle BCAA utilization is indirect via its lipolytic action on adipose
tissue
o First step in catabolism of BCAA is transamination with -KG to form glutamate and transamination of
glutamate with pyruvate to form alanine (glucogenic amino acid)
o Alanine is released from fasting muscle to be converted to glucose in the liver and kidney

 Metabolic fuel during prolonged of fasting = ketone bodies (acetoacetate + beta-hydroxybutyrate)

o These are synthesized by the liver
o Usage of these ketone bodies spares glucose for tissues that utilize glucose as their primary source of
fuel e.g. brain
o CPT deficiency  accumulation of lipid in muscle cells, can’t transport fatty acids across the IMM to be
oxidized for energy
- TETANIC CONTRACTION
 Total amount of glycogen in skeletal muscle exceeds amount of glycogen in the liver
 Even through the amount of glycogen per unit weight in muscle is approx. 10% of that found in the liver, since
skeletal muscle mass is over 25x greater, more glycogen is present
 The glycogen in muscle is used as an energy source during tetanic contraction + anoxia
 Muscle glycogen, in contrast to liver glycogen, isn’t depleted in fasting and isn’t broken down to glucose for
use by other tissues because muscle doesn’t have the enzyme glucose 6-phosphatase (used in
gluconeogenesis, liver and kidneys)
 But there are 2 different systems have been found for the activation of the phosphorylase in muscle
1. Epinephrine induced phosphorylation  activates the phosphorylase system in muscle, but time
required is about 3 min (slow)
2. Contraction induced phosphorylation  activates the phosphorylase system by the stimulatory action
of Ca2+ on both activated and nonactivated phosphorylase kinase

Purine nucleotide cycle:

 Metabolic pathway used to:
o Regulate the levels of adenine nucleotides
o Formation of NH4+ in contracting muscle
o Fumarate production to replenish it in the TCA cycle
o Oxidation of malate to OAA to replenish NADH needed in respiratory chain
 o Controls level of adenine nucleotides, Pi and NH4+ which are regulators of glycolysis 2 the PFK step
 Specifically for skeletal muscle it also helps dispose of the AMP produced by the reaction catalyzed by myokinase
 Sequence of 3 enzymatic reactions
 Reaction
 Enzymes involved: AMP deaminase, adenylosuccinate synthetase, adenylosuccinase, glutamate-oxaloacetate
aminotransferase, fumarase, malate dehydrogenase

6. Chemical composition and metabolic features of red and white fibers of the striated muscle
- There are at least 2 types of striated muscle fibers (red and white) in skeletal muscle and others are intermediates
between the 2
- RED FIBERS VS WHITE FIBERS
 Red fibers have large amount of sarcoplasm with more nuclei, mitochondria and mitochondrial iron-containing
cytochromes
 Red fibers have more myoglobin  gives it the red color
 More lipid droplets
 Red fibers also twitch longer, contract slower and are more easily tetanized than white fibers
- 3 types of skeletal muscle fibers have been seen based on their metabolic + physiological characteristics:
1. Fast-twitch white fibers: low respiration, highly glycogenolytic, high myosin ATPase activity
2. Fast-twitch red fibers: high respiration, high glycogenolytic, high myosin ATPase activity
3. Slow-twitch red fibers: high respiration, low glycogenolytic, low myosin ATPase activity

7. Cardiac muscle. Regulation of cardiac muscle contraction

- CARDIAC MUSCLE
 Cardiac muscle is also striated like skeletal muscle
 Has a more irregular shape and contains branching fibers
 Same contractile proteins are present (actin, myosin, etc.) and perform the same functions
 Cardiac SR also contains at least 4 of the major proteins found in the SR of skeletal muscle (ATPase, LMW
proteolipid + 2 Ca binding proteins)
- REGULATION OF CARDIAC MUSCLE CONTRACTION
 Force and rate of contraction of the heart are stimulated by EPI
 Cardiac contraction process:
o EPI stimulates cAMP formation
o cAMP-dependent protein kinase is stimulated  catalyzes the phosphorylation of:
 Myofibrillar proteins TN-I and myosin light chain
 SR membrane protein
 Phospholamban (regulates active Ca2+ transport in cardiac SR)
- Unphosphorylated state  phospholamban is an inhibitor of cardiac muscle
sarcoplasmic reticulum Ca2+-ATPase which transports calcium from cytosol into the
sarcoplasmic reticulum
- When phosphorylated (by PKA)  disinhibition of Ca2+-ATPase of SR leads to faster
Ca2+ uptake into the sarcoplasmic reticulum
 Sarcolemma membrane proteins
o Ca2+ concentration in the sarcoplasm is increased by entry through the sarcolemma + release from SR
o Entry of Ca2+ through the sarcolemma is accompanied by an efflux of Na  forms a Ca-Na exchange
system  3 Na ions per one Ca ion
o Negatively charged phospholipids e.g. phosphatidylinositol and phosphatidylserine are involved in the
exchange process
o After contraction, Ca is sequestered back into the SR

8. Metabolic fuel of cardiac muscle

- Lots of different sources for metabolic fuel for the heart
- 60-90% from fatty acid oxidation
- AFTER A MEAL
 Glucose, pyruvate, lactate (since levels of free fatty acids are low)
 Both pyruvate and lactate inhibit the uptake and oxidation of FFA and
BCAA
- FASTING
 Short-term fasting  FFA (triacylglycerols) + BCAAs
 o Fatty acids mobilized from adipose tissue via action of lipoprotein lipase which hydrolyzes plasma
triacylglycerols
o The FFA that enter the cells have to migrate through the cytoplasm to reach the OM where the fatty
acids are activated into acyl CoA which is facilitated by a protein
o BCAAs are also used
o The RLS of the use of BCAAs by the heart is -keto acid dehydrogenase
 Uses as its substrates the -keto acids from leucine, isoleucine and valine
 Pyruvate can inhibit usage of BCAAs by inhibiting this enzyme
 Long-term fasting  ketone bodies
 Fatty acids and ketone bodies inhibit usage of glucose and pyruvate by inhibiting glucose entry into the cell and
inhibiting the enzyme PFK
- ANOXIA
 Glucose + glycogen
 Glucose utilization is the same as in skeletal muscle
 3 mechanisms have been suggested for stimulation of the breakdown of heart glycogen:
1. EPI activation of glycogen phosphorylase (1st enzyme of glycogenolysis)
2. Inhibition of phosphorylase phosphatase by AMP formed during anoxia
3. Activation of phosphorylase b by AMP
 Supplies of cardiac glycogen are replenished after anoxia by activation of glycogen synthase (converts UDP-
glucose to UDP) which gets activated (DP) by a specific phosphatase
- ALCOHOL INGESTION  acetate
- Increase in mechanical load by the heart is accompanied by:
 Increase in O2 consumption
 Increase in oxidative phosphorylation
 Shift from glucose to palmitate (fatty acid) usage
o The carnitine acyltransferase system helps couple the rate of FA uptake by cardiac mitochondria to TCA
cycle activity
o Hence, the rate of translocation of acyl units across the IMM limits the rate of long-chain fatty acyl
carnitine oxidation

9. Distribution and chemical composition of adipose tissue

- ADIPOSE TISSUE
 Adipose tissue is a specialized mammalian tissue and has an important role in energy metabolism
 After a meal, end products of the digestion of dietary fat, carbohydrates and protein are converted to
triacylglycerols by the adipose tissue
 During fasting, adipose tissue triacylglycerols are hydrolyzed to glycerol and fatty acids and the fatty acids can
be used by a variety of other tissues as their metabolic fuel e.g. skeletal muscle and cardiac muscle
 Triacylglycerols are both a major store of metabolic fuel AND a source of energy for heat production
 Oxidation of fatty acids yields 2x more energy than carbohydrates or proteins per unit weight
 The deposition of triacylglycerols in specialized cells eliminates the need for extensive storage of carbohydrate
or triacylglycerol in other tissues
- DISTRIBUTION + CHEMICAL COMPOSITION
 Widely distributed  muscles, under skin, around BV,
abdominal cavity
 2 types of adipose tissue = white and brown
o White adipose tissue
 Energy storage
 Spherical cells that contain:
- Large vacuole of triacylglycerol that
occupies almost the entire cell (80% of
white adipose tissue, 99% of lipid content)
- Mitochondria along the periphery of the central lipid droplet
- Flattened Golgi near the nucleus
- ER
 Approx. 20 fatty acids are found in adipose triacylglycerol (e.g. oleic acid, palmitic acid, linoleic
acid, stearic acid, myristic acid)
o Brown adipose tissue
 Heat production
 Polygonal cells that contain:
- A more abundant cytoplasm than white adipose cells
- Small lipid droplets
- Nucleus isn’t flattened
- Small amounts of ER and Golgi
- Lots of mitochondria
 Both types of adipose tissue are well supplied with blood capillaries and are well innervated
10. Adipose tissue metabolism after feeding and in fasting
- FED STATE
 Triacylglycerols
o Triacylglycerols in the diet are digested in the stomach and small intestine by lipases
o Principal products are 2-monoacylglycerols and free fatty acids
o These get absorbed by the small intestine’s epithelial cells which assemble the monoacylglycerols and
free fatty acids into triacylglycerols  get packaged into chylomicrons
o Also in the fed state, the liver synthesizes free fatty acids from excess carbohydrate and amino acids
o These fatty acids are assembled into triacylglycerols and packaged into VLDLs
o The triacylglycerols in chylomicrons and VLDLs are hydrolyzed to fatty acids and glycerol by lipoprotein
lipase (LPL)
 This enzyme is located on the surface of capillary endothelial cells in adipose tissue (+skeletal
muscle)
 Activation of LPL requires: insulin and phospholipids
 Insulin stimulates the rate of LPL synthesis and its secretion from the adipocyte to its
endothelial site of activity
 ApoC-II found in the chylomicrons and VLDL activates hydrolysis by binding the lipoproteins to
the enzyme
 Products of the reaction are: free fatty acids and glycerol
- Free fatty acids  enter the adipocyte with the help of a protein transporter and is
esteried to form triacylglycerols, allows deposition of fuel
- Glycerol  transported through the bloodstream and taken up by the liver to be used
in glycolysis or gluconeogenesis
o Lipoprotein particles remaining after action of LPL on VLDL are LDLs
 Glucose
o Most of the glucose taken up by adipose tissue after a meal is used in triacylglycerol synthesis, but
small amounts are converted to glycogen (somehow involves insulin)
o The glycogen formed in adipose tissue after feeding is eventually later converted to triacylglycerols as
well
o Glucose entry into adipocyte
 Entry of glucose into the adipocyte is the RDS and occurs by a process of passive mediated
transport aided by 2 membrane glycoproteins
 Compounds with sulfhydryl groups inhibit adipocyte glucose transport activity and if we were
to treat adipocytes with these compounds, the ability of insulin to stimulate adipocyte glucose
transport will be eliminated
 A number of oxidants (e.g. H2O2) also stimulate glucose transport in the adipocyte and insulin
has been found to stimulate H2O2 production hence increasing glucose transport
 Insulin also may induce glucose transport via GLUT4 through rapid and reversible translocation
of glucose transport proteins from a large intracellular pool to the plasma membrane
 Hence stimulation of glucose transport into the adipocyte by insulin is of great importance
o Once inside the adipocyte, glucose is rapidly phosphorylated by hexokinase to form glucose-6-
phosphate and is under the control of insulin and/or glucose itself (GLYCOLYSIS)
 Adipose tissue in the fed state has higher hexokinase activities leading to GLYCOLYSIS
 But, a portion (23%) of glucose taken up by the adipocyte is metabolized via HMP shunt and the
remainder by glycolysis
 Insulin stimulates both glucose transport into the adipocyte and the enzyme hexokinase =
increase in glycolytic rate
o The stimulation of these 2 pathways increases production of NADH and NADPH
 Some NADH is used for reduction of DHAP to G3P
 Some NADH is transhydrogenated to form NADPH  this reaction is necessary in adipose tissue
because the HMP shunt produces only 65% of the NADPH needed for fatty acid synthesis
o Pyruvate produced by glycolysis is converted to glycerol and fatty acid moieties of the triacylglycerols
that are synthesized in adipose tissue
 Pyruvate is also converted to OAA and acetyl CoA and then to citrate which leaves the mitochondria to be
converted back into acetyl CoA and OAA in the citrate cleavage pathway
o Transfer of citrate out means that there’s a loss of OAA from mitochondria
o Mitochondria are impermeable to cystolic OAA and therefore to replace mitochondrial OAA, the OAA is
converted to malate, then to pyruvate which can be transported across the mitochondrial membrane
o Pyruvate is then carboxylated via pyruvate carboxylase in the mitochondrial matrix
 Increases in the rate of glycolysis and HMP shunt after feeding are accompanied by increases in the activites of:
o Pyruvate dehydrogenase
o Acetyl CoA carboxylase
o Citrate cleavage enzyme (ATP citrate lyase)
o Fatty acid synthetase
 Glucose can also be converted directly into palmitic acid (fatty acid) via acetyl CoA without involvement of the
entire TCA cycle and involves:
o Glycolysis
 o HMP shunt
o Transhydrogenation
o Pyruvate  OAA, acetyl CoA and citrate
o Citrate cleavage to OAA and acetyl CoA
o Acetyl CoA  palmitate via lipogenesis
 Adipose tissue will remove amino acids from the blood for conversion into fatty acids and proteins
o Insulin stimulates transport of amino acids into the adipocytes and directly affects adipocyte protein
synthesis
o The amino acids differ in their ability to form fatty acids, leucine has the greatest ability since the end
products of its metabolism are acetoacetate and acetyl CoA
o Acetoacetate will become incorporated into fatty acids in adipose tissue
- FASTED STATE
 During fasting, the triacylglycerols stored in adipose tissue are used as fuel
 Triacylglycerols in adipose tissue are hydrolyzed to glycerol and free fatty acids by hormone sensitive lipase
(HSL)
 HSL is located within adipocytes
 HSL is activated by: glucagon, EPI, ACTH, GH, thyroxine, secretin and glucocorticoids
o Glucagon and EPI are the principal lipolytic hormones after long-term fasting/starvation
o These hormones (except glucocorticoids) activate adenylate cyclase in the adipocyte membrane
o Increases cAMP levels
o Activates protein kinase which phosphorylates the lipase = active form of HSL is its phosphorylated
form
o EPI and glucocorticoid effect on HSL doesn’t involve cAMP
o Products = monoacylglycerols and free fatty acids
 Insulin and glucose inhibit fatty acid mobilization from adipose tissue, and since the levels of these 2
compounds are present @ low levels during fasting, mobilization of fatty acids from adipose tissue occurs
 Prostaglandins also have an antilipolytic effect by reducing stimulation of lipolysis by the hormones AND
inhibiting cAMP formation

11. Triacylglycerol synthesis in adipose tissue. Role of adipose tissue in heat production
- TRIACYLGLYCEROL SYNTHESIS
 Fatty acids derived from dietary triacylglycerol OR from glucose and amino acids are used to synthesize
triacylglycerols in adipose tissue
 3 pathways of triacylglycerol synthesis:
o Glycerol-3-phosphate pathway (MAJOR)
o Monoacylglycerols pathway
o DHAP pathway
 Glycerol-3-phosphate pathway
o Glycerol-3-phosphate can come from:
 Reduction of DHAP (most tissues)
 In fed state  the DHAP is derived from glucose via glycolysis
 In fasting state  glyceroneogenesis (adipose tissue and liver)
 Also, in the liver glycerol can be phosphorylated by glycerol kinase
o The acyl CoA:sn-glycerol 3-phosphate acyltransferase that catalyzes the acylation of glycerol-3-
phosphate is increased after feeding
o When the triacylglycerol is synthesized it is composed of 3 fatty acids attached to a glycerol
 @ sn1 = saturated fatty acids
 @ sn2 = unsaturated fatty acids
 @ sn3 = more random, with preference for LCFA
o The acyltransferase enzyme has specificity with respect to the acyl CoA used in the esterification
o A protein “specifier factor” has been found that interacts with acyltransferases to direct the acylation
of palmitate to position 2
- HEAT PRODUCTION
 Heat production by brown adipose tissue  occurs primarily in newborns and adult hibernating animals
 Heat production by white adipose tissue  in adults during exposure to low temperatures
 Heat production is started by EPI
 Activates hydrolysis of adipose tissue triacylglycerols to fatty acids and glycerol
 Released fatty acids have a dual role in heat production:
o Source of reducing equivalents (via beta oxidation that produces FADH2, NADH) for energy production
by respiratory chain
o Act as uncouplers of OP
 Therefore, energy produced by ETC in the respiratory chain of enzymes is released as heat instead of being
converted to ATP
 Uncoupling is attributed to fatty acids acting as carrier molecules for protons into the mitochondria =
eliminates the proton gradient and bypasses ATPase involved in generation of ATP
 Liver basics:
- Structural unit of the liver = lobule containing parenchymal cells/hepatocytes
- The liver has a dual afferent blood supply:
 Portal vein  carries blood from the GI tract, spleen and pancreas containing all the digested nutrients and
toxins
 Hepatic artery
- Blood from the branches of the 2 vessels mix when passing through the sinusoidal capillaries of the liver lobules
- The sinusoids drain into the central veins of the lobules which are branches of hepatic veins
- Branches of the afferent vessels with the bile ducts form the portal triad and run along the edges of each lobule
- Sinusoidal capillaries pass through the parenchyma from the periphery of the lobule to the central vein
- Small bile capillaries run between the parenchymal cells, anastomose at the periphery of the lobule and enter the bile
ducts
- The sinusoid walls are lined by Kupffer cells (functional phagocytes, large cells with bulging nuclei)

12. Carbohydrate metabolism in liver

- LIVER FUNCTIONS
 Provides metabolic fuel for a variety of tissues in fasting e.g. glucose, amino acids, ketone bodies
 Remove metabolic waste products generated by other tissues
 Synthesize structural compounds for other tissues
 Detoxification
- CARBOHYDRATE METABOLISM
 Major function of the liver is blood glucose homeostasis
o After a meal  liver converts glucose in blood to glycogen (GLYCOGENESIS)
o In fasting  liver converts glycogen to glucose (GLYCOGENOLYSIS) AND also converts amino acids and
lactate into glucose
 In the fed state, the uptake of glucose by hepatocytes (in contrast to adipose/muscle cells) isn’t dependent on
insulin, uses GLUT2 that isn’t insulin dependent
 BUT, insulin does simulate glucose metabolism by the liver by stimulating glucokinase activity (glucokinase is in
liver, also known as hexokinase isozyme IV, vs. hexokinase normally)
o Glucose will get converted to glucose-6-phosphate
o Most of the glucose-6-phosphate is converted to glycogen via GLYCOGENESIS
o Small amounts of glucose-6-phosphate enter the glycolytic pathway and HMP shunt for fatty acid
synthesis
 HMP shunt produces NADPH and ribose-5-phosphate
 NADPH  reducing agents, fatty acid synthesis, cholesterol metabolism, nucleotide synthesis,
NT synthesis
 Ribose-5-phosphate  DNA/RNA synthesis, ATP synthesis, NAD+ synthesis, FAD+ synthesis,
CoA synthesis
 During fasting, glycogen is broken down to glucose-1-phosphate via glycogen phosphorylase during
GLYCOGENOLYSIS (under control of glucagon and EPI)
o Effects of EPI
 EPI is released into the blood from chromaffin cells of the adrenal medulla
 Mediated by -adrenergic receptors and involve a cAMP-independent mechanism AND -
adrenergic receptors that do involve cAMP production
 -adrenergic receptors  increases Ca levels, activates phosphorylase kinase = activates
glycogen phosphorylase = GLYCOGENOLYSIS
o Effect of glucagon
 Released from alpha cells of the pancreas
 Mediated by -adrenergic receptors that do involve cAMP production
 Adenylate cyclase is stimulated and activates glycogen phosphorylase = GLYCOGENOLYSIS
 BUT, the amount of glycogen in the liver isn’t really sufficient enough to maintain blood glucose levels during
fasting, especially 12h after a meal
o Hence, the major source of blood glucose in prolonged fasting is GLULCONEOGENESIS in the liver (also
stimulated by glucagon and epinephrine)
o Gluconeogenesis is stimulated by the glucocorticoids in prolonged fasting and is inhibited by insulin
after a meal
 Approximately 15% of the glucose metabolized by the liver is via the HMP shunt
o Generates NADPH for fatty acid and cholesterol synthesis
o Glucose metabolism by HMP shunt is increased in feeding and decreased in fasting
o After feeding, glucose-6-phosphate dehydrogenase increases 5x
 The amount of glucose metabolized by GLYCOLYSIS in liver is small when compared to other tissues e.g. muscle,
brain
o Main purpose of glycolysis in the liver is to furnish pyruvate for conversion to acetyl CoA that will be
used in fatty acid synthesis
o Rate of glycolysis in liver is increased in feeding and decreasing in fasting (similar to the HMP shunt)
13. Lipid and protein metabolism in liver. The metabolic fuel of liver
- LIPID METABOLISM
 Fed state  liver synthesizes fatty acids from glucose and incorporates them into triacylglycerols and
phospholipids
 Fate of fatty acids synthesized by the liver = VLDL which can then be deposited as triacylglycerols
 Fasted state  liver will use free fatty acids released from adipose tissue as its metabolic fuel
 The carnitine transport system that is used for entry of fatty acids into the mitochondria is the RDS/regulates
usage of fatty acids
 Hence in the fasting state, fatty acid oxidation in the mitochondrial matrix occurs
 And in the fed state, incorporation of fatty acids into phospholipids occurs
- PROTEIN METABOLISM
 Both protein synthesis and breakdown occurs in the liver
 The ONLY serum proteins not synthesized in the liver are the gamma globulins
 Plasma proteins are constantly removed from circulation and pinocytosis into hepatocytes
 They are then hydrolyzed to amino acids which are then used for extrahepatic tissues
 Half-life of both serum and liver protein = 10 days
 Half-life of muscle protein = 180 days
- METABOLIC FUEL OF LIVER
 Energy demands of liver are large
 After a meal  supplied by lactate
 During fasting  fatty acids released from adipose tissue
o Liver does have LPL activity therefore plasma triacylglycerols are also a possible source of fatty acids for
the organ in fasting

Liver biotransformation:
- Lots of biotransformative reactions occur solely in the liver
- Examples:
 Synthesis of bile acids from cholesterol
 Reduction of adrenal steroids + conjugation with glucuronic acid or sulfuric acid
 Conjugation of androgens and estrogens with glucuronic acid or sulfuric acid
 Formation of bile pigments
 Synthesis of purine and pyrimidine bases
 Formation of uric acid
 Urea synthesis
- The liver detoxifies a variety of drugs and poisons via oxidation, reduction, hydrolysis, conjugation and methylation

14. Biotransformation reactions in liver: oxidative, reductive and hydrolytic reactions

- OXIDATION
 Enzymes that catalyze oxidation reactions are present within the microsomal fractions of liver
o Microsomes are fragments of ER and attached ribosomes obtained by centrifugation of liver cells
o Rough microsomes are dedicated to protein synthesis
o Smooth microsomes are rich in enzymes responsible for oxidative reaction, especially the
MFOs/monooxygenases
o Activity of these enzymes requires:
 Reducing agent (NADPH)
 Molecular O2
o Catalyze oxidative reactions commonly called hydroxylation reactions (which includes aromatic and
aliphatic hydroxylation, N-, O- and S-dealkylations, sulfoxidation, epoxidation)
 The microsomal oxidase systems/MFOs/monooxygenase system has 3 components and is referred to as the
P450 system:
1. Cytochrome P450
2. Flavoprotein reductase
3. Phospholipid
 Examples:
o Oxidation of toluene  p-cresol
o Oxidation of barbiturates (shouldn’t be taken with alcohol and medication), polycyclic hydrocarbons
o Oxidation of alcohols  aldehydes or ketones AND aldehydes  carboxylic acids
 Catalyzed by 2 groups of enzymes in liver that do not involve the cytochrome P450 system:
1. Pyridine nucleotide linked oxidoreductases
- Use NAD+
- Located in both cytosol + mitochondria
- e.g. liver alcohol dehydrogenase  catalyzes the oxidation of a number of
aldehydes (formaldehyde, acetaldehyde) to their acids
2. Aldehyde oxidase
- Metalloflavoprotein containing iron, molybdenum and FAD
 - Catalyzes oxidation of benzaldehyde to benzoic acid
- REDUCTION
 Hepatic microsomal enzymes reduce both azo and nitro compounds via addition of H
 Enzymes involved: azoreductase and nitroreductase
 Azoreductase + nitroreductase activity are due to both NADPH-cytochrome P450 reductase and cytochrome
P450
 Examples:
o Reduction of nitrobenzene  aniline
o Reduction of aldehydes and ketones  alcohols (by liver aldehyde/ketone reductases and NADPH
coenzyme)
- HYDROLYSIS
 Requires water and can be catalyzed by enzymes e.g. esterases, hydrases
 Found in plasma, brain, intestinal mucosa, erythrocytes and muscle + liver
 Esterase is found in human liver that catalyzes the hydrolysis of:
o Acetanilide  aniline + acetic acid
o Procaine
o Xylocaine
o Alipathic esters
 Epoxide hydrase catalyzes the conversion of epoxides to diols e.g. cyclohexane epoxide  cyclohexane diol

15. Biotransformation reactions in liver: conjugation, methylation and other detoxication

reactions
- CONJUGATION
 Glucuronic acid
o 4 types of compounds are conjugated with glucuronic acid:
1. Hydroxyl (both phenolic and alcoholic)
2. Carboxyl
3. Sulfhydryl
4. Amino
o Enzyme: UDP-glucuronyltransferase (ER)
o Co-factor = UDP-glucuronic acid
o Major pathway of drug metabolism
 Sulfate
o Compounds with alcoholic or phenolic hydroxyl groups undergo conjugation with sulfate
o Enzyme: sulfotransferase (cytosol of liver cell)
o Sulfate group comes from 3’-phosphoadenosine 5’-phosphosulfate (PAPS)
 Glutathione
o Glutathione conjugates are thioethers
o These thioethers formed by combination of alkyl and aryl halides, epoxides and alkenes with
glutathione
o Enzyme: glutathione transferases (cytosol)
o Some glutathione conjugates are converted to mercapturic acids catalyzed by S and N-
acetyltransferases
o Glutamine
o Conjugated with phenylacetic acid and drugs
o Example  phenylacetyl CoA + glutamine forming phenacetylglutamine and CoASH
o Enzyme: acyl CoA:amino acid N-acyltransferase
o Conjugation with acetic acid detoxifies sulfanilamide
o Conjugation of benzoic acid with glycine forms hippuric acid (also by acyl CoA:amino acid N-acyltransferase like
glutamine)
- METHYLATION
 Enzyme: methyltransferases (liver)
 Use S-adenosylmethionine as methyl donor
 Example: catechol-O-methyltransferase catalyzes the O-methylation of NE, dopamine, dopa and drugs
 Example: methylation of H2S to form methylmercaptan

- OTHER DETOX REACTIONS

 Cysteine conjugate of aromatic drugs  sulfhydryl derivative + NH3 + acetic acid (cysteine conjugate -lyase)
 Arylhydroxamic acids  arylhydroxylamines (arylhydroxamic acid acyltransferase)
 Arylhydroxylamine  N-acyloxyarylamine
 Disulfides sulfhydryl compounds
o Catalyzed by thioltransferases
o Transfer of thiol residue to glutathione
  2-oxyaldehydes  -hydroxy acids
o Glyoxylase system consists of 2 enzymes: glyoxylase I and II
o Intermediate formed is a thioester of glutathione
 Cyanide is detoxified by sodium thiosulfate  sodium thiocyanate + sodium sulfate (rhodanese)
- Overall idea: products of detox reactions mostly are usually less toxic than the reactants, but this isn’t always the case

16. Metabolic fuel of kidney

- Uses 7 sources: palmitate, lactate, citrate, glucose, glycerol, glutamine, ketone bodies
 The choice of source depends on availability of the substrates
 Palmitate is the main metabolic fuel for the kidney (60-80%)
 Lactate is in second place
 In the presence of palmitate, lactate usage is inhibited BUT the uptake of lactate is not
 Lactate is taken up by the kidney and is converted to glucose via GLUCONEOGENESIS
 Uses of citrate, glycerol and glutamine haven’t yet been defined
 Ketone bodies are used during long-term fasting/starvation AND diabetes
 Glucose usage is small, but is generally used after feeding and during anoxia
- Source of energy for ATP production is via aerobic oxidation of substrates, glycolysis produces 4% of the required ATP
- ATP is required for reabsorption of NaCl by the kidney tubules
- KIDNEY CORTEX + MEDULLA METABOLISM
 Metabolism in different parts of the kidney differ
 Kidney cortex  outer part of kidney, contains glomerulus + convoluted tubules
o High respiratory metabolism:
 High rate of oxygen consumption
 Lots of TCA cycle and respiratory chain enzymes
o Fatty acids are the main source of metabolic fuel and after fatty acid oxidation ketone bodies are
formed (like the liver)
o Oxidation of palmitate, lactate, glucose and ketone bodies to be used as metabolic fuel mainly occurs
in the kidney cortex
 Medulla  innermost part of the kidney, split into pyramidal sections, contains nephrons
o Low respiratory metabolism:
 Lots of glycolytic enzymes
 Low energy requirement (from glycolysis mainly)
 Low rates of oxygen consumption
 Low amounts of TCA cycle and respiratory chain enzymes
o The source of glucose for glycolysis carried out by the kidney medulla is plasma glucose after a meal OR
glucose synthesized in the cortex during fasting

17. Ammonia production in kidney and gluconeogenesis in kidney cortex

- GLUCONEOGENSIS IN KIDNEY CORTEX
 The synthesis of glucose in the kidney cortex via gluconeogenesis is much faster than that in the liver
 BUT, glucose synthesized by the kidney cortex doesn’t really go into the bloodstream
 Instead, it has been suggested that the glucose goes to the kidney medulla where there are lots of glycolytic
enzymes
 Rate of gluconeogenesis is affected by:
o EPI + glucagon (like in the liver, activates the process via cAMP)
o Nutritional state (faster in fasted state)  during the fasting state, ketone bodies and fatty acids supply
the energy needed for kidney cortex gluconeogenesis
o Acidosis (unclear)
 Gluconeogenesis is heavily regulated @ the PEPCK step
 PEPCK can be induced during acidosis
 Accompanied by increases in the amount of enzyme itself + increases in the amount of mRNA
for the enzyme (therefore affects the synthesis of the enzyme)
- AMMONIA PRODUCTION
 Production and excretion of ammonia is the kidney’s response to acidosis
 In acidosis, lots of H+ ions are present in excess in the blood and by producing ammonia the kidney
avoids/protects the situation from escalating
 Renal ammoniagenesis is initiated by a phosphate dependent glutaminase
 Phosphorylates glutamine to form glutamic acid and NH4+
 This enzyme is located in the inner membrane/matrix of mitochondria
 The formation of ammonia during acidosis is due to increases in the amount or activity of glutaminase OR
increased usage of its feedback inhibitor glutamate (hence the enzyme is no longer inhibited)
 Ammonia production could also be by the pyrimidine nucleotide cycle
- KIDNEY TUBULAR TRANSPORT MECHANISMS
  The kidney nephron is specialized for blood filtration and urine excretion
 Nephron is composed of various segments: glomerulus, proximal convoluted tubule, descending and ascending
loops of Henle, distal convoluted tubule and the collecting duct
 The nephron filters and then selectively reabsorbs various compounds e.g. water, NaCl, potassium,
bicarbonate, glucose, amino acids
 The nephron tubules are lined with cells that facilitate absorption from the lumen to the interstitium/blood
o These cells are polarized
o Consists of 2 membrane sides  apical membrane (faces lumen/urine) and basolateral membrane
(faces interstitium/blood)
 The enzyme Na+-K+ ATPase is present within these cells especially @ high concentrations within the proximal
convoluted tubules and ascending loop of Henle
 Function: pumping 3 Na outside, 2 K inside via usage of ATP (therefore there’s a 2:3 ratio)
 Proximal convoluted tubules of the nephron reabsorb the most substances (like the first checkpoint)
o 100% reabsorption of glucose and amino acids (cotransported with Na+)
 If a patient develops high glucose concentration in the blood e.g. diabetes, glucose will start to
appear (approx. 160 mg/dl)
 If it further rises to 350 mg/dl all the transporters will become saturated leading to glucose in
the urine (diabetes mellitus)
 In Hartnup disease there’s no tryptophan transporter in the proximal tubule leading to
tryptophan deficiency
 Tryptophan is a precursor for niacin, therefore niacin deficiency also develops leading to
pellagra (plaques + desquamation)
o 67% reabsorption of water, bicarbonate, K, P and NaCl
o NaCl reabsorption also occurs using ATP derived from aerobic oxidation of substrates + some glycolysis
o The reabsorption of Na here is driven by the Na+-K+ ATPase pump found in the basolateral side
o The pump is responsible for the functioning of the proximal convoluted tubule and consumes lots of
ATP
o What it does:
 Creates a low Na concentration inside the cells of the proximal convoluted tubule
 The low Na concentration drives reabsorption other solutes and water into the blood e.g.
glucose and Na are cotransported
- Obviously under normal circumstances the proximal tubules reabsorb all glucose
- Once there’s low concentration of Na within the cell, the high Na concentration in the
lumen/urine facilitates development of a concentration gradient to which Na can
diffuse down coupled with glucose
 Creates a high K concentration of potassium inside the cell
 The concentration gradient of K drives transport of K back to the outside but linked to chloride
which is also reabsorbed
 Then the low concentration of Cl left behind in the cell can drive absorption of Cl coupled with
entry of anions into the lumen/urine e.g. OH, formate, sulfate
o Finally, once there’s a high concentration of Na outside the cell via osmosis water can be reabsorbed
back into the blood
 Ascending loop of Henle
o Reabsorption of NaCl
o Again uses the Na+-K+ ATPase using ATP derived from aerobic oxidation of substrates, some glycolysis
o NaCl will get reabsorbed from the lumen of the ascending loop of Henle into the peritubular space

Nervous tissue basics:

- Neuron = functional unit of the nerve cell, 10 billion in the NS
- Has lots of cytoplasmic processes  neuron contact with other neurons, glands, muscles
- Composed of 3 main elements:
1. Cell body/perikaryon/soma
 Central large nucleus
 Abundant cytoplasm
2. Axon
 Long extension of the cytoplasm that conducts generated impulses away from the cytoplasm
 In the CNS, axons are present in white matter
 In the PNS, axons are in nerve bundles
3. Dendrites
 Short extensions of the cytoplasm that surround the cell body
 Receive impulses from other neurons
- Cytoplasm contents of the cell body and cytoplasmic extensions are different
 Cell body  Golgi, mitochondria, pigment granules (melanin, lipofuscin), lipid droplets, glycogen droplets,
neurofibrils, neurofilaments, microfilaments and RER referred to as Nissl bodies
 Axoplasm  neurofibrils and mitochondria, no lipid droplets or glycogen droplets and there’s absence of the
RER (indicates the proteins are synthesized within the cell body and are transported down to the axons and
dendrites via anterograde transport)
- Protected by structures e.g. myelin produced by OD in the CNS and Schwann cells in the PNS
 - Gray matter contains the cell bodies of neurons, dendrites + axon proximal portions
- White matter consists of myelin and axons

18. Nervous tissue lipids and proteins

- LIPIDS
 Nervous tissue is composed of a high content of lipids with unique structures
 Main metabolic role of lipids in nervous tissue is for cell membrane function as well as a component of myelin,
white and gray matter
 The most abundant substance in the brain is water
 The second most abundant substance in the brain is cholesterol and about 25% of the body’s cholesterol is
present in nervous tissue
 Cholesterol has a low rate of breakdown and low turnover and is synthesized within the brain
 Fatty acid contents:
o Most of the fatty acids in the brain are synthesized in situ
o Only small amounts of fatty acids are incorporated from the diet
o Fatty acid content of lipids are quite constant but can be altered depending on the lipid composition of
the diet
o Fatty acid synthesis in the brain of course involves the initial step of palmitate synthesis via malonyl
CoA intermediate, but is elongated to form fatty acids with 22-24 carbons and the fatty acids are also
unsaturated
 Elongation occurs by both the mitochondrial and microsomal/ER systems
- Microsomal elongation is occurs via a similar process to fatty acid synthesis  malonyl
CoA is the source of 2 carbon units, NADP provides reducing power
- Mitochondrial elongation  uses acetyl CoA, NADPH,NADH and is almost a reversal of
beta oxidation where enoyl CoA reductase replace the acyl-CoA dehydrogenase in beta
oxidation
 Desaturation also occurs via 2 enzymatic systems that desaturate between 6-7 and 9-10
carbons
 Lipids found in nervous tissue are present in different concentrations at different locations:
o Cholesterol + sphingomyelin (ceramide phosphocholine)  equal in myelin, WM, GM
 Sphingomyelin in myelin  lignoceric and nervonic acid
 Sphingomyelin in gray matter  stearic acid
 Excessive accumulation of sphingomyelin occurs in Niemann-Pick disease
o Phosphatidylinositol  myelin
o Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine  GM
 Source of choline + serine  exogenous (from blood)
 Source of ethanolamine + inositol  endogenous
o Gangliosides  GM (Tay-Sachs)
o Sulfatides + cerebrosides  myelin and WM (metachromatic leukodystrophy)
- PROTEINS
 3 major proteins in nervous tissue
1. S-100
o Highly acidic
o Found in large amounts in glial cells, small amounts in neurons
o Glutamic + aspartic acids are 30% of amino acid content
o Has 3 non-identical subunits
o High affinity for Ca and undergoes a conformational change when Ca is present
2. 14-3-2
o Named based on its behavior in chromatography
o Highly acidic
o Found in neurons
o Isozyme of enolase (catalyzes reversible conversion between 2-PG and PEP during glycolysis)
3. Calmodulin
o Ca binding protein
o Found in brain
o Needed for Ca dependent activation of cyclic
nucleotide phosphodiesterase
o Also needed for Ca dependent release of Ach
and norepinephrine from vesicles
 Both S-100 and 14-3-2 have been implicated in learning + memory
 2 major proteins in myelin:
1. Basic protein
o 30% of total protein in myelin
o Large number of arginine residues
2. Proteolipid
o Combination of protein with lots of hydrophobic amino acids + lipid
  Basic protein + proteolipid are located on the outer and inner surfaces of the myelin membrane and abut each
other in the lipid bilayer
 Neurotubules, neurofilaments and microfilaments
o These transverse the cytoplasm from one dendrite to another OR dendrite to axon
o Role in axonal flow
o Microtubules
o Involved in regulation of cholesterol synthesis in glial cells
o Involved in cell differentiation, IC transmission of signals from CSR and effectors of enzyme
transport
o Major protein = tubulin (14% protein in brain, 2 similar subunits)
o Neurofilaments  protein subunit distinct from tubulin
o Neurofibrils are bundles of neurofilaments
o Microfilament protein = actin
 Rapid turnover rate and products are reutilized unlike other tissues that release breakdown products for liver
processing
o Half-life of brain proteins is 20x faster than total body protein
o This constant amount of protein in brain, high proteolysis, rapid incorporation of glucose into proteins
and small A-V differences of nitrogenous compounds for the brain  indicates retention + reutilization
of NH4+ by nervous tissue
o Some urea cycle enzymes are present for this but are unknown why
o Retention + reutilization of phospholipids also occurs (only tissue that does this)
o This retention of proteins + lipids are logical since the BBB is impermeable and promotes retention
AND during starvation the brain is resistant to deterioration

- ACTION POTENTIAL
 Consists of 3 stages: resting, depolarization and repolarization
 The resting membrane potential is about -60 mV (neuron is more negative on the inside
 There is a high ratio of potassium ions inside to outside
 There is a low ratio of sodium ions outside to inside
 The ratios of these two ions is maintained by the Na+-K+ ATPase which pumps 3 Na out for every 2 K in
 When a stimulus arrives, the membrane suddenly becomes permeable to Na ions allowing Na ions to enter
allowing the inside of the cell to become more positive = depolarization (+35 mV)
 The resting membrane potential is restored during repolarization which is where the membrane becomes
permeable to K+ ions which leaks outside, accompanied by less influx of Na
 Even though the resting membrane potential is restored, the appropriate concentrations of K and Na are not
present within the cell
 The appropriate gradient is restored by the Na+-K+ ATPase which requires energy
 Like skeletal muscle and cardiac muscle, the brain has a reservoir of high energy phosphate in the form of
phosphocreatine

19. Carbohydrate metabolism in nervous tissue

- Fed state  glucose
 Glucose is used as the only metabolic fuel during the fed state
 Completely oxidized to CO2 and H20
 25% of O2 consumption is attributed to glucose oxidation in the brain of adults (vs 50% in infants)
 Glycolysis occurs at 20% capacity
 TCA cycle is at full capacity
 Small amounts of lactate leave the brain
- Anoxia  glycolysis and lactate formation
 Anoxia is the biggest threat to the survival of the brain
 But the brain is protected by the Pasteur effect
 Rates of both glycolysis and lactate formation are also increased  attributed to activation of hexokinase +
PFK-1
 PFK-1 is also activated by NH4+ which is found in increased levels during anoxia
- Fasting state  glycogen (small amount of glucose supply), ketone bodies
 Endogenous glycogen can only supply a small amount of glucose in the absence of blood glucose
 If there is a sudden removal of glucose from the blood, cerebral failure will occur
 BUT, if there is gradual removal/long-term hypoglycemia like in starvation cerebral failure doesn’t occur
 Instead ketone bodies replace glucose as metabolic fuel
- Synthesis and breakdown of brain glycogen
 Both the D and I forms of glycogen synthase are found in brain along with the enzymes that catalyze their
interconversions
 Insulin doesn’t have an effect (can’t cross BBB)
- 3-5% of glucose metabolized in the brain is via HMP shunt (replenishes NADPH for fatty acid and cholesterol synthesis)

20. Amino Acid metabolism in nervous tissue

21. The Eye. Metabolic features of lens and cornea
22. Metabolic features of retina. Role of rhodopsin in vision

23. Factors that regulate production of red and white blood cells
24. Structure and metabolic features of red blood cells
25. Glucose-6-phosphate deficiency and methemoglobinemia, hereditary spherocytosis and
elliptocytosis
26. Blood group systems. H, A and B substances. The Rh factors and the MNSs blood group
system
27. Metabolic features of neutrophils
28. Activation of neutrophils. Myeloperoxidase and proteinases of neutrophils