Acidic dissociation
1. General expression: HA is the acid (proton donor) and A- is the conjugate base (proton
acceptor):
2. An acid dissociates in water to yield a hydrogen ion (H+) and its conjugate base
3. A base combines with H+ in water to form its conjugate acid
k1[HA] = forward rate, k-1[H+][A-] = reverse rate
Measures of acidity
1. pKa
When forward & reverse rates are equal, acidic dissociation constant, Ka, is defined by:
o k1/k-1 = [H+][A-] / [HA] = Ka
Expresses the STRENGTH OF AN ACID
pKa = -log[Ka]
Strong acid has pKa of W2 (H+ binds loosely to conjugate base)
Weak acid has a pKa of D10 (H+ binds tightly to conjugate base)
2. pH
Henderson-Hasselbalch equation: pH = pKa + log [A-]/[HA]
A measure of the ACIDITY OF A SOLUTION
pH = -log[H+]
Neutral solution has a [H+] of 10-7 pH = 7
Acidic solution has a [H+] > 10-7 pH < 7
Alkaline solution has a [H+] < 10-7 pH > 7
Buffers and Buffering capacity
1. A solution that contains a mixture of a weak acid and its conjugate base
2. It resists changes in [H+] on addition of acid or alkali
3. The buffering capacity of a solution is determined by the acid-base concentration and the pKa
of the weak acid
Maximum buffering effect occurs when:
o [weak acid] = [conjugate base]
When the buffer effect is at its maximum:
Lysine
Arginine
Histidine
6. Acidic, polar
Aspartic acid
Asparagine
Glutamic acid
Glutamine
Secondary structures of proteins and collagen
1. Secondary structure = arrangement of H bonds between peptide nitrogens & carbonyl oxygens
of different amino acids
2. Helical coils
Hydrogen-bonded nitrogens & oxygens are on nearby amino acids
Right-handed alpha helix most common
o Alpha-keratin in hair and nails
o Myoglobin has several alpha-helical regions
Proline, glycine, and asparagine helix breakers
3. Beta sheets (pleated sheets) may run parallel or antiparallel
Hydrogen bonds between residues on neighboring peptide chains
4. Left-handed helical strands
Wound into a supercoiled triple helix in collagen
Collagen major structural protein in the body
o Primary structure: repeating glycine-X-Y sequences
o X and Y are freqeuntly proline or lysine
o Contains hydroxyproline & hydroxylysine
Protein Denaturation Agents
1. Extremes of pH (e.g., strong acid or alkali)
2. Ionic detergents (e.g., sodium dodecylsulfate/SDS)
3. Chaotropic agents (e.g., urea, guanidine)
4. Heavy metal ions (e.g., Hg++)
5. Organic solvents (e.g., alcohol or acetone)
6. High temperature
7. Surface films (e.g., as when egg whites are beaten)
Sickle cell anemia
1. Caused by mutant sickle cell hemoglobin (Hgb S)
Hydrophobic valine replaces hydrophilic glutamate at position 6 of the beta-chain of normal
hemoglobin A (Hgb A)
2. Sickle cell disease
Individuals with homozygous genotype (SS)
Have only Hgb S in their RBCs
Symptoms
o Severe anemia: deoxy Hgb S produces fibrous precipitates formation of sickle cells shorter
life span severe anemia
o Acute episodes of vaso-occlusion sickle cell crisis
Sickle cells cant pass thru capillaries vasocclusion
Disabling pain that requires hospitalization
3. Sickle cell trait
Individuals with heterozygous genotype (AS)
Have both Hgb A and Hgb S in their RBCs
Symptoms
o Usually asymptomatic
o Episodes of hematuria
Scurvy
1. Defective collagen synthesis resulting from a vitamin C (ascorbic acid) deficiency
2. Consequences of abnormal collagen
Defective wound healing
Defective tooth formation
Loosening of teeth
Bleeding gums
Rupture of capillaries
3. Ascorbic acid is required for hydroxylation of proline and lysine during post-translational
modification of collagen
4. Pathogenesis
Hydroxylating rxn requires hydroxylase, O2, & Fe2+
Vit C is required to maintain iron in its oxidation state (Fe2+)
Hydroxyproline forms H-bonds that stabilize collagen helix
Symptoms of scurvy are thus the result of weakend collagen when these hydrogen bonds are
missing
Free energy change
1. The quanitity of energy from chemical reactions that is available to do work (DG)
2. The DG of a rxn A + B C + D is:
DG = DG0 + RTln ([C][D]) / [A][B])
o where DG0 is the standard free-energy change (when concentrations of all reactants and
products are 1M and pH = 7), R is the gas constant (1.987 cal/molK) and T is the absolute
temperature
3. When the rxn has reached equilibrium:
DG0 = RTlnKeq
Thermodynamic spontaneity: Exergonic and Endergonic Rxns
1. Exergonic rxns are spontaneous (rxn goes to the right)
Keq > 1
DG0 is negative
Final concentration of the products, C and D, is greater than that of the reactants, A and B
2. Endergonic rxns are nonspontaneous (rxn goes to the left)
Keq < 1
DG0 is positive
Final concentration of the reactants, A and B, is greater than that of the products, C and D
3. Note: DG0 CANNOT predict spontaneity under intracellular conditions
Intracellular spontaneity is a function of actual concentrations as well as Keq, DG, NOT DG0
Enthalpy, entropy, and free-energy change
1. Enthalpy (DH)
The amount of heat generated or absorbed in a rxn
2. Entropy (DS)
Measure of the change in randomness or disorder of system
when a salt crystal dissolves, when a solute diffuses from a more concentrated to a less
concentrated solution, and when a protein is denatured
when a complex molecule is synthesized from smaller substrates
3. Free-energy change (DG)
Is related to enthalpy and entropy as follows:
o DG = DH - T DS (where T = absolute temp in Kelvins)
Catalysts and the Rate of Reaction
1. Rate of reaction
The DG0 provides no info concerning the rate of conversion from A to B
When A is converted to B, it first goes through an energy barrier called the transition state, AB
The activation energy (DG) = energy required to scale the energy barrier and form the
transition state
The greater the DG, the lower the rate of the rxn converting A to B
2. Catalysts (mostly enzymes) result in a:
Lower DG
Faster reaction rate
Michaelis-Menten equation
1. Describes the kinetics of enzyme rxns
2. In enzyme-catalyzed rxns:
k1 k3
E + S ES E + P
k2
Where E=enzyme, S=substrate, P=product, k1-3 = rate constants
3. Velocity (v) of product formation is related to [ES]:
o v = k3[ES] where k3 is also called kcat
4. Michaelis-Menten eq predicts velocity if [enzyme] is constant:
Where Vm = max velocity & Km is the Michaelis constant
5. Km = [substrate] at which v = Vm ([S] = Km)
6. A plot of velocity versus [S] is a rectangular hyperbola
Lineweaver-Burk Equation
1. Form of the Michaelis-Menten eq, which is sometimes known as the double-reciprocal
equation:
1 = Km + [S] = Km x 1 + 1
v = Vm[S] Vm [S] Vm
2. The slope is Km/Vm
3. The Y-intercept = 1/Vm
4. The X-intercept is 1/Km
Enzyme Regulation:
How doe Inhibitors affect the Lineweaver-Burk plots?
1. Competitive inhibitors
apparent Km
Vm remains the same
slope
X-intercept has smaller absolute value
Y-intercept is unchanged
2. Noncompetitive inhibitors
Vm
Km unchanged
slope
X-intercept is unchanged
Y-intercept is larger
3. Uncompetitive inhibitors (bind only to ES complex)
Both Km & Vm are different lines on the plot are parallel
Allosteric regulation of enyzmes: Definition, How do they affect Km, and Example of
Hexokinase
1. Low-molec wgt effector binds to enzyme at a specific site other than active site (the allosteric
site) & alters its activity
2. Allosteric enzymes usually have >1 subunit and >1 active site
Active sites that interact cooperatively: velocity vs, [S] = sigmoid
Binding of 1 substrate facilitates binding of substrate at other sites
3. Effectors may have a + or effect on activity
Positive effectors the apparent Km
Negative effectors the apparaent Km
4. Example: muscle hexokinase
Hexokinase catalyzes 1st rxn in use of glucose my muscle cells:
o Glucose + ATP glucose-6-P + ADP
Hexokinase has a low Km compared to blood [glucose], so it is saturated and operates at its Vm
When glycolysis slows down, gluc-6-P accumulates gluc-6-P allosterically inhibits
hexokinase (keeps gluc-6-P in balance)
Other mechanisms of enzyme regulation:
1. Induction/repression of enzyme synthesis
2. Covalent modificataion
3. Protein-protein interaction
1. Induction/repression of enzyme synthesis
Cytochrome P450 enzymes in the liver (degrade and detoxify drugs) are induced by the drugs
themselves
2. Covalent modificataion
Phosphorylase (enzyme that breaks down glycogen) is activated by phosphorylation of a
specific hydroxyl group
This phosphorylation is stimulated by hormones that elevate blood glucose, such as glucagon
and Epi
3. Protein-protein interaction between enzyme & regulatory protein
Pancreatic lipase (digests dietary fat) is assisted by colipase
Colipase anchors lipase to the surface of fat droplets
Mechanism and Treatment of Methanol & Ethylene glycol Poisoning
1. Mechanism of poisoning
Toxicity is caused by the action of their metabolites
In both cases, the 1st oxidation is carried out by alcohol dehydrogenase
o Methanol formaldehyde + formic acid
Eyes very sensitive to formaldehyde blindness
1. Uncoupling
Carry H+ across inner mit membrane w/o going thru complex V
This short-circuits the proton gradient and uncouples electron flow from ATP synthesis
Energy, instead of used to make ATP, is dissipated as heat
Uncoupling agents:
o Dinitrophenol (2,4-DNP) former diet drug
Caused blindness (retina has rate of oxidativ metblism)
o Thermogenin helps to maintain normal body temp
Found normally in brown fat of newborn mammals
2. Inhibitors (via blocking e- flow thru complexes or direct action)
Complex I Amobarbital (barbiturate), Rotenone (insecticide), Piericidin A (antibiotic), Amytal
Complex II Antimycin A (antibiotic)
Complex IV Cyanide, Hydrogen sulfide, Carbon monoxide
ATP synthase Oligomycin
Carbohydrate digestion and absorption
1. Digestion
Disacharides (sucrose),oligosacharides (dextrins),& polysacharides (starch) are cleaved into
monosaccharides (glucose, fructose)
Starch: storage from of carbs in plants
o Hydrolyzed to maltose, maltotriose, and a-dextrins by a-amylase in saliva and pancreatic juice
Disaccharides & oligosaccharides
o Hydrolyzed to monosaccharides by enzymes on the surface of epithelial cells in the small
intestine
2. Absorption
Monosaccharides absorbed directly by carrier-mediated transport
These sugars (primarily glucose) travel via portal vein to liver for:
o Oxidation to CO2 and H2O for energy
o Storage as glycogen
o Conversion to triglyceride (fat)
o Release into general circulation (as glucose)
Glycogen metabolism
[Glycogen: carb storage, found chiefly in liver & muscle]
1. Glycogenesis (glycogen synthesis)
Activated substrate: Uridine diphosphate-glucose
Glycogen synthase adds to nonreduc end of chains in a-1,4 links
Branching enzyme amylo (14) to (16) transglycosylase creates branches w/a-1,6 linkages
Stimulator: insulin (via dephosphorylation in muscle, liver, & fat)
Inhibitors: glucagon (liver), Epi (muscle & liver), phosphorylase (liver), cAMP, Ca2+ (muscle)
3. The liver releases the resynthesized glucose into the circulation for transport back to the
muscle
Regulation of glycolysis
1. All are the irreversible steps:
Fructose 6-P fructose-1,6-BP via phosphofructokinase
o Stimulators: AMP, fructose 2,6-BP (in liver)
o Inhibitors: ATP, citrate
o Rate-limiting step
D-glucose glucose-6-P via hexokinase/glucokinase*
o Inhibitors: glucose-6-P
PEP pyruvate via pyruvate kinase
o Inhibitors: ATP, alanine
o Stimulators: fructose-1,6-BP (in muscle)
Pyruvate acetyl CoA via pyruvate dehydrogenase
o Stimulators: CoA, NAD, ADP, pyruvate
o Inhibitors: ATP, NADH, acetyl CoA
2. Induced by insulin
Gluconeogenesis regulation
1. All are the irreversible steps:
Pyruvate OAA via Pyruvate carboxylase (mitochond)
o Requires biotin, ATP
o Activated by acetyl CoA
OAA PEP via PEP carboxykinase
o Requires GTP
Fructose-1,6-BP fructose-6-P via Fructose-1,6-BPase
Glucose-6-P glucose via Glucose-6-phosphatase
2. These enzymes are only found in liver, kidney, intestinal epithel
3. Muscle cannot participate in gluconeogenesis
4. Hypoglycemia is caused by a deficiency of these key enzymes
5. Induced by glucocorticoids, glucagon, cAMP
6. Suppressed by insulin
Pnemonic: Pathway Produces Fresh Glucose
Pyruvate dehydrogenase complex
1. Contains 3 enzymes that require 5 cofactors:
Pyrophosphate (from thiamine)
Lipoic acid
CoA (from pantothenate)
FAD (riboflavin)
NAD (niacin)
2. Reaction:
Pyruvate + NAD+ + CoA acetyl-CoA + CO2 + NADH
3. The complex is similar to the a-ketoglutarate dehydrogenase complex (same cofactors, similar
substrate and action)
4. Cofactors are the first 4 B vitamins plus lipoic acid:
B1 (thiamine; TPP)
B2 (FAD)
B3 (NAD)
B5 (pantothenate CoA)
Lipoic acid
Pentose phosphate pathway
1. Sites: lactating mammary glands, liver, adrenal cortex all sites of fatty acid or steroid
synthesis
2. Begins with glucose 6-P
3. The irreversible oxidative portion generates NADPH
NADPH needed for: fatty acid and cholesterol (steroid) synthesis, maintaining reduced
glutathione inside RBCs
4. The reversible nonxidative portion rearranges the sugars so they can reenter the glycolytic
pathway
5. Ribose 5-P, which is needed for nucleotide synthesis, can be formed from glucose 6-P by
either arm
6. Major regulatory enzyme: glucose 6-P dehydrogenase
Glucose 6-P 6-phosphogluconolactone
7. Stimulators: NADP+, insulin
8. Inhibitors: NADPH
Sucrose and Lactose Metabolism
1. Sucrase converts sucrose to glucose and fructose
Hexokinase can convert fructose fructose 6-P (muscle, kidney)
Fructose enters glycolysis by a different route in the liver
Dihydroxyacetone phophate (DHAP) enters glycolysis directly
After glyceraldehyde is reduced to glycerol, it is phosphorylated and then reoxidized to DHAP
2. Lactase converts lactose to glucose + galactose
Galactokinase converts galactose galactose 1-P
Galactose 1-P glucose 1-P glycolysis
Lipoprotein absorption
1. Exogenous lipid (from intestine), except for medium-chain fatty acids, is released into the
plasma as chylomicrons
Chylomicrons contain a high proportion of TGs
TG is hydrolyzed to FFAs and glycerol by lipoprotein lipase on the surface of capillary
endothelium in muscle and adipose tissue
The cholesterol rich chylomicron remnants travel to the liver, where they are taken up by
receptor-mediated endocytosis (RME)
2. Endogenous lipid (from liver) is released into blood as VLDLs
VLDL TG is hydrolyzed by lipoprotein lipase to FFAs and glycerol, yielding low-density
lipoproteins (LDLs)
LDLs are removed from circulation by RME in tissues that contain LDL receptors (tissues that
need cholesterol, but mostly in liver)
LDL cholesterol:
o Inhibits HMG CoA reductase (RLS in cholesterol synthesis)
o Down-regulates LDL receptor synthesis LDL uptake
High density lipoproteins (HDLs) are made in the liver
Lipoprotein functions and associated apolipoproteins:
Chylomicrons
1. Delivers dietary triglycerides to peripheral tissues and dietary cholesterol to liver
2. Secreted by intestinal epithelial cells
3. Excess causes pancreatitis, lipemia retinalis, eruptive xanthomas
4. Associated apolipoproteins:
B-48 mediates secretion
As are used for formation of new HDL
C-II activates lipoprotein lipase
E mediates remnant uptake by liver
Lipoprotein functions and associated apolipoproteins:
VLDL
1. Delivers hepatic triglycerides to peripheral tissues
2. Secreted by liver
3. Excess causes pancreatitis
4. Associated apolipoproteins:
B-100 mediates secretion
C-II activates lipoprotein lipase
E mediates remnant uptake by liver
Ketogenesis
1. The formation of acetoacetate and b-hydroxybutyrate from metabolism of acetyl CoA in the
liver
2. Reaction:
Acetyl CoA + acetoacetyl CoA hydroxymethylglutaryl CoA (HMG CoA)
HMG CoA acetoacetate and acetyl CoA
Acetoacetate b-hydroxybutyrate (requires NADH)
3. How is acetoacetate used in the body?
Extrahepatic tissues (especially heart) can activate acetoacetate at the expense of succinyl CoA
and burn acetoacetyl CoA for energy
Glucose-starved brain can use acetoacetate for fuel (b/c its freely soluble in blood and easily
crosses the BBB)
Fatty acid synthesis
1. Carried out by fatty acid synthase, a cytosolic complex
2. Primary substrates:
Acetyl CoA: formed in mitoch, mainly by pyruvate dehydrogenase
o Its transported to cytosol by citrate-malate-pyruvate shuttle
Malonyl CoA: formed by biotin-linked carboxylation of actyl CoA
3. The acetyl and malonyl moieties are transferred from the sulfur of CoA to activate sulfhydryl
groups in the fatty acid synthase
4. 7 cycles lead to production of palmityl:enzyme, which is hydrolyzed to yield products
palmitate & fatty acid synthase
5. Palmitate is the precursor for longer & unsaturated fatty acids
Chain-lengthening occurs in the mitoch and ER (C16C18etc)
Desaturating system is also present in the ER
o Requires NADPH and O2
o Inserts double bonds no further than 9 carbons from the carboxylic acid group
What do the limitations of the desaturating system result in?
1. The limitations of the desaturating system impose a dietary requirement for essential fatty
acids (those w/double bonds >10 carbons from the carboxyl end)
Lineoleic acid
o Precursor for arachidonic acid (which is beginning of cascade that synthesizes prostaglandins,
thromboxanes, and eicosanoids)
Linolenic acids
Glycerolipid synthesis
1. This process is carried out by the liver, adipose tissue, and the intestine
2. Pathways begin w/glycerol 3-P, which is mainly produced by reducing dihydroxyacetone
phosphate w/NADPH
3. Succesive transfers of acyl groups from acyl CoA to carbons 1 and 2 of glycerol 3-phosphate
produce phosphatidate, which can then be converted to a variety of lipids:
Triacylglycerol (from transfer of acyl group from acyl CoA)
Phosphatidyl choline & phosphatidyl ethanolamine (from transfer of base from its cytidine
diP/CDP derivative)
Phosphatidylserine (from exchange of serine for choline)
Phosphatidylinositol (from reaction of CDP-diacylglycerol with inositol)
Sphingolipid synthesis
1. Begins with palmityl CoA and serine
Produces dihydrosphingosine and sphingosine
2. Sphingosine can then by acylated to produce ceramide
Additional groups may be added to the C1-OH of ceramides
Cholesterol synthesis
1. Cholesterol is made by the liver and intestinal mucosa from acetyl CoA in a multistep process
2. Key intermediate = HMG CoA
HMG CoA reductase: regulatory enzyme that catalyzes HMG CoA + NADPH mevalonic acid
Increasing amounts of intracellular cholesterol lead to inhibition of HMG CoA reducate and
accelerated degradation of the enzyme
3. Overall reaction:
Acetoacetyl CoA + acetyl CoA HMG CoA synthase HMG CoA HMG CoA reductase
mevalonic acid cholesterol
What are the fates of the products of cholesterol synthesis?
1. Mevalonic acid
Precursor of a number of natural products called terpenes, which include vit A, vit K, coenzyme
Q, and natural rubber
2. Cholesterol
Converted to steroid hormones in the adrenal cortex, ovary, placenta, and testes
Majority is oxidized to bile acids in the liver
7-dehydrocholesterol is the starting point for synthesis of vit D
Lipid malabsorption
1. May be caused by insufficent removal of NH4+, resulting from disorders that involve one of
the enzymes in the urea cycle
2. Signs and Symptoms
Blood NH4+ concentrations above the normal range (30-60 mM)
Mental retardation, seizure, coma, and death
3. Enzyme defects
Low activity of carbamoyl P synthetase or ornithine-carbamoyl transferase [NH4+] in blood
& urine NH4+ intoxication
Defective argininosuccinate synthetase, argininosuccinase, OR arginase blood levels of
metabolite preceding defect
o NH4+ levels may also rise
4. Treatment
Restriction of dietary protein
Intake of mixes of keto acids that correspond to essential amin acid
Eating benzoate & phenylacetate: alternate path for NH4+ excretion
Carbon skeletons of amino acids
1. Amino acids can be grouped into families based on the point where their carbon skeletons
enter the TCA cycle
2. AcetylCoA/Ketogenic fam(blue:keto-& glucogenic; red:ketogen only)
Isoleucine, leucine, lysine, phenylalanine, tryptophan, and tyrosine
Phenylalanine tyrosine via phenylalanine hydroxylase
Tyrosine is starting compound for:
o Epi and NE, T3 and T4, Dopamine, Melanin
3. a-Ketoglutarat fam (arginine,histidine,glutamate,gluatmine,proline)
Histidine precursor of histamine
Glutamate excitatory neurotransm, can be converted to GABA
4. Succinyl CoA family (isoleucine, methionine, valine)
Methyl of methionine participates in methylation rxns as S-adenosylmethionine (SAM)
5. Fumarate family (phenylalanine and tyrosine)
6. Oxaloacetate family (asparagine and aspartate)
7. Pyruvate fam (alanine,cysteine,glycine,serine,threonine, tryptophan)
Essential amino acids
1. Isoleucine
2. Leucine
3. Lysine
4. Phenylalanine
5. Tryptophan
6. Histidine
7. Methionine
8. Valine
9. Threonine
Phenylketonuria (PKU)
1. Results from a deficiency of:
Phenylalanine (Phe) hydroxylase OR
Dihydropteridine reductase
2. Findings
Phe in the blood (hyperphenylalaninemia)
Phe builds up to toxic concentrations in body fluids, resulting in CNS damage with mental
retardation
Phe inhibits melanin synthesis hypopigmentation
3. An alternative pathway for Phe breakdown produces phenylketones, which spill into th eurine
4. In those affected, tyrosine is an essential amino acid
5. Treatment: restricting dietary protein (phenylalanine)
Albinism
1. No tyrosinase (1st enzyme on pathway to melanin)
2. Have little or no melanin and are:
Easily sunburned
Very susceptible to skin carcinoma
Photophobic b/c of lack of pigment in iris of eye
Homocystinuria
1. May result from several defects:
Cystathionine synthase (CS) deficiency
affinity of CS for its coenzyme, pyridoxal phosphate (PLP) (may respond to megadoses of
pyridoxine/vit B6)
Methyl tetrahydrofolate homocyst methyltransferase deficiency
Vit B12 coenzyme deficiency (may respond to vit B12)
2. Finding: homocysteine accumulation in blood, appears in urine
3. Pathologic changes
Dislocation of optic lens
Mental retardation
Osteoporosis and other skeletal abnormalities
Atherosclerosis and thromboembolism
4. Pts unresponsive to vitamin therapy may be treated with:
GMP synthesis
Balances supply of adenine and guanine ribonucleotides
4. Interconversion among purines ensures control of their levels
AMP deaminase converts AMP back to IMP
GMP reductase converts GMP back to IMP
IMP is the starting point for synthesis of AMP and GMP
Origin of the atoms in the pyrimidine ring
De novo pyrimidine synthesis
1. In mammals, 1st 3 steps occur on one multifunctional enzyme called CAD, which stands for
the names of the enzymes
CO2 + glutamine CAP synthetase II carbamoyl-P (CAP)
Synthesis of dihydroorotic acid is a 2-step process:
o Committed step: aspartate + CAP aspartate transcarbamoylase carbamoyl aspartate
o Carbam aspartate dihydrorotase dihydroorotic acid + H2O
2. Dihydroorate forms UMP
Dihydroorate orotic acid
Orotic acid + PRPP orotidylate (OMP)
Decarboxylation of OMP uridylate (UMP)
These 2 steps occur on 1 protein (if defected: orotic aciduria)
3. Synthesis of remaining pyrimidines involves UMP
Phosphorylation of UMP UDP + UTP
Addition of amino group from glutamine to UTP CTP
Regulation of pyrimidine synthesis
1. CAP synthetase II regulation
Inhibited by UTP
Activated by ATP and PRPP
2. CTP itself inhibits CTP synthetase
Salvage of pyrimidines
Accomplished by the enzyme pyrimidine phosphoribosyl transferase, which can use orotic acid,
uracil or thymine, but NOT CYTOSINE
Deoxyribonucleotide synthesis
Formation of deoxyribonucleotides (for DNA synthesis) involves reduction of sugar of
ribonucleoside diphosphates:
1. Ribonucleotide reductase
deoxyribonucleotides)
2. Aminopterin and methotrexate
Inhibit dihydrofolate reductase (enzyme that converts dihydrofolate to tetrahydrofolate)
3. Fluoredeoxyuridylate
Inhibits thymidylate synthetase (enzyme that converts dUMP to dTMP)
Gout
1. May result from a disorder in purine metabolism
2. Is associated w/hyperuricemia
3. Primary gout: overproduction of purine nucleotides
Mutations in PRPP synthetase loss of feedback inhibition by purine nucleotides
A partial HGPRT deficiency less PRPP is consumed by salvage enzyes PRPP activates
PRPP amidotransferase
4. Secondary gout
Due to radation therapy, CA chemo (b/c they cell death)
5. Treatment: allopurinol
Xanthine oxidase catalyzes oxidation of allopurinol to alloxanthine, which is a potent inhibitor
of the enzyme
Result: uric acid levels fall, hypoxanthine & xanthine levels rise (is OK, b/c they dont form
crystals)
Energy expenditure (3 components)
1. Basal energy expenditure (BEE)
resting energy expenditure
Energy used for metabolic processes at rest
Represents >60% of total energy expenditure
Related to the lean body mass
2. Thermic effect of food
Energy required for digesting and absorbing food
Amounts to ~10% of energy expenditure
3. Activity-related expenditure
20-30% of daily energy expenditure
Caloric yield from foods and what % they should be in diet
1. Carbs: 4 kcal/g
50-60% of caloric intake
2. Proteins: 4 kcal/g
10-20% of caloric intake (0.8 g/kg body weight/day)
3. Fats: 9 kcal/g
No more than 30% of caloric intake
4. Alcohol: 7 kcal/g
Fats:
Essential fatty acids, Deficiency, and Excess storage
1. Essential fatty acids (EFAs):
Linoleic acid
Linolenic acid
2. Deficiency
Mainly seen in low-birth-weight infants maintained on artificial formulas and adults on TPN
Characteristic system: scaly dermatitis
3. Excess fat
Stored as triacylglycerol
Marasmus vs. Kwashiorker
Marasmus Kwashiorker
Insufficient food, including both calories and protein Starvation with edema often due to protein
deficient diet
Depleted subQ fat Pitting edemaFlaky paint dermatosis: dark patches on skin that peel
Liver ketogenesisbrain&heart fuel Large liver due to fatty infilatration
Muscle wasting (break protein for gluconeogenesis & protein synthes) Muscle wasting less
severe
Frequent infections Frequent infections
Low body temp
Micronutrient deficiencies Other nutrient deficiencies
Slowed growth(<60% expected wgt) Growth failure(>60% expected wgt)
Death when energy & protein reserves exhausted Poor appetitie (anorexia)Watery stools
w/undigested foodMental changes (apathetic)
Vitamin A
1. Functions:
11-cis-retinal prosthetic group of rhodopsin
Beta-carotene antioxidant NOT TOXIC at high doses
Retinyl phosphate mannose acceptor/donor in glycoprotein synth
Retinol & retinoic acid regulate tissue growth & differentiation
2. Deficiency signs and symptoms:
Night blindness, Xerophthalmia (cornea keratinizes: Bitot spots)
o Muscle weakness
o Hemolytic anemia (b/c of fragility of RBCs)
o Retinal degeneration
Vitamin K
1. Function
Post-translational carboxylation of glutamyl residues in Ca+-binding proteins: factors VII, IX,
&X
2. Deficiency ( PT, aPTT, but nl bleeding time)
Impaired blood clotting bruising, bleeding
Causes:
o Fat malabsorption
o Drugs that interfere w/vit K metabolism (warfarin)
o Antibiotics that suppress bowel flora
3. Vitamin K in infants
Neonates are born w/low stores of vit K
Vit K crosses placental barrier poorly
Newborns given single injection of vit K
High doses: anemia, hyperbilirubinemia, kernicterus
The B vitamins
1. B1 = Thiamine
2. B2 = Riboflavin
3. B3 = Niacin
4. B5 = Pantothenate (pantothenic acid)
5. B6 = Pyridoxine (pyridoxamine, pyridoxal)
6. B12 = cobalamin
Thiamine (vitamin B1)
1. Thiamine pyrophosphate (TPP): required for nerve transmission & is coenzyme for several
key enzymes:
Pyruvate & a-ketoglutarate dehydrogenase(glycolysis, TCA)
Transketolase (pentose phosphate pathway)
Branched-chain keto-acid dehydrogenase (valine, leucine, isoleucine metabolism)
2. Deficiency leads to beriberi, which occurs in 3 stages:
Early: loss of appetite, constipation, nausea, periph neuropathy, irritability, fatigue
Moderately severe: Wernicke-Korsakoff syndrome (mental confusion, ataxia, ophthalmoplegia)
Severe (in addtion to polyneuritis):
3. Clinical usefulness:
High doses: tx homocystinuria (defective cystathione b-synthase)
Vitamin B6: Pantothenic acid
1. Function
Essential component of coenzyme A (CoA) and of fatty acid synthase
Cofactor for acyl transfers
2. Deficiency (very rare)
Vague presentation, little concern to humans
Dermatitis, enteritis, alopecia, adrenal insufficiency
Biotin
1. Function
Covalently linked biotin = prosthetic group for carboxylation enzymes (e.g. pyruvate
carboxylase, acetyl CoA carboxylase) (NOT decarboxylations)
2. Deficiency (rare)
Signs and symptoms:
o Dermatitis
o Hair loss
o Atrophy tongue papilla
o Gray mucous memb
o Paresthesa,muscle pain
o Hypercholesterlemia
o ECG abnormalities
Causes
o Antibiotic use (since intestinal bacteria make biotin)
o Eating Avidin (raw egg whites)
Binds biotin in a nondigestible form
If you eat >20 eggs/day
Folic acid
1. Function
Polyglutamate derivatives of tetrahydrofolate serve as coenzymes in 1-carbon transfer rxns:
o Purine & pyrimidine synthesis
o Thymidylate synthesis
o Conversion of homocysteine to methionine
o Serine-glycine interconversion
2. Deficiency
Signs & symptoms:
o Megaloblastic anemia
o Neural tube defects
o blood homocysteine associated w/atherosclerotic disease
Can be caused by several drugs:
o Methotrexate (chemo)
o Trimethoprim (antibact)
o Pyrimethamin(antimalari)
o Diphenylhydantoin (anticonvulsant)
o Primidone (anticonvuls)
Vitamin B12 (cobalamin)
1. Functions
Coenzyme for methylmalonyl CoA succinyl CoA (methylmalonyl CoA mutase) in propionyl
CoA metabolism
Coenzyme for methyl transfer between tetrahydrofolate & methionine (homocysteine methyl
transferase)
2. Deficiency:
Signs & Symptoms:
o Megloblastic anemia
o Paresthesia, optic neuropathy, subacute combined degenerat
o Prolonged deficiency irreversible nervous system damage
Causes:
o Intake of no animal products (vegans)
o Achlorhydria, intrinsic factor (impaired absorption)
o Malabsorption (impaired pancreatic function, sprue, enteritis, D. latum, absence of terminal
ileum/Crohns)
3. Use Schilling test to detect deficiency
Vitamin C (ascorbic acid)
1. Functions
Coenzyme for re-dox rxns, including:
o Post-translational hydroxylation of proline & lysine in maturation of collagen
o Carnitine synthesis
o Tyrosine metabolism
o Catecholamine neurotransmitter synthesis
Antioxidant
Facilitator of iron absorption
2. Deficiency
Signs & symptoms:
o Mild: capillary fragility w/easy bruising & petechiae (pinpoint hemorrhages in skin), immune
function
o Severe: scurvy ( wound healing, osteoporosis, hemorrhage, anemia, swollen gums, teeth may
fall out)
y
Symptoms of Mineral Deficiencies
Mineral Deficiency-Associated Conditions
Calcium ParesthesiaTetanyBone fractures, bone painOsteomalacia (as in vit D deficiency)
Iodine GoiterCretinism
Iron AnemiaFatigue, tachycardia, dyspnea
Magnesium Neuromusc excitability, paresthesiaDepressed PTH release
Phosphorus (as phosphate) Deficiency rarely occurs
Zinc Growth retardation & hypogonadismDry, scaly skinMental lethargyImparied taste & smell,
poor appetite