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Course Objective Review the biochemical processes most frequently tested on the NBDE
enz (z) A B
Michaelis-Menton Kinetics
[substrate] vs. Vo is hyperbolic At high [substrate] the reaction reaches its maximum velocity (Vmax) The Km is the [substrate] at which the reaction velocity is Vmax The Km is an effective measure of affinity.
Enzymes with a low Km have a high affinity for substrate Enzymes with a high Km have a low affinity for substrate
Hexokinase vs Glucokinase
Hexokinase- Ubiquitously expressed High affinity for glucose Low Km and Vmax Saturated at low [glucose] Glucokinase- Liver specific enzyme High Km and Vmax Saturated at high [glucose]
Hexokinase functions during fasting Glucokinase functions after a glucose rich meal
of a
Enzyme Inhibition
Competitive vs. Noncompetitive Competitive Inhibitors bind the same site on the enzyme as the substrate increase the Km of the enzyme do not affect the Vmax
Competitive Inhibitors
Vmax is unchanged in the presence of a competitive inhibitor
Km is increased
Vmax is unchanged
Enzyme Inhibition
Competitive vs. Noncompetitive Noncompetitive Inhibitors do not bind the same site as the substrate do not affect the Km of the enzyme decreases the Vmax
The reaction will never reach the original maximum velocity, regardless of the substrate concentration
Noncompetitive Inhibitors
Vmax is decreased in the presence of noncompetitive inhibitor
Km is unchanged
Vmax is decreased
Part II figure-3
Carbohydrates
General formula (CH2O)n Functions: Immediate Energy Source Blood Glucose Energy Stores Extracellular Matrix Glycoconjugates Nucleic Acid Precursor Glycogen (homopolymer) Glycosaminoglycans (heteropolymer) Glycoproteins Glycolipids Ribose
Carbohydrates
Disaccharides Lactose = Sucrose = Maltose = Polysaccharides Starchamylose (14 linkage) amylopectin, branched (14, 16) polysaccharides of glucose (16, 13, and 14) glucose + galactose glucose + fructose glucose + glucose
Monosaccharides
Can be in an open chain form or a cyclic form In the open chain form, each has a carbonyl.
Six carbon sugars tend to be cyclic Cyclization generates an anomeric carbon isomers, and Lehninger Figure 7-6
two
Disaccharides
Lactose = galactose(14) glucose Anomeric carbon for galactose is C-1 Anomeric carbon for glucose is C-1 C-1 of glucose remains free Lactose is a reducing sugar Reducing End of Lactose
Sucrose = fructose
glucose(12)
Anomeric carbon for glucose is C-1 Anomeric carbon for fructose is C-2 No free anomeric carbon Sucrose is not a reducing sugar Lehninger Figure 7-12
Starch
Storage form of glucose in plants Is made of amylose and amylopectin Amylose polymerized glucose no branches (14) linkage
(14)
Amylopectin polymerized glucose branched (14) linkage (16) at branch points many reducing ends for quick mobilization
Glycogen
Primary linkage (14)
Storage form of glucose in humans Primarily stored in liver functions to maintain blood glucose levels Stored in muscle functions to provide glucose during exercise
Heteropolysaccharides Glycosaminoglycans
GAGs are long, unbranched heteropolysaccharide chains composed of repeating disaccharide units. Modified by negatively charged carboxyl and sulfate groups Extended, unbranched chains surrounded by a shell of water gel-like matrix or ground substance Neighboring molecules tend to repel each other slippery nature of mucous secretions and synovial fluid Compressible but rebound after the release of pressure resilience of synovial fluid and vitreous humor GAGs vary based on their disaccharide subunits, linkage, sulfation patterns and distribution in the human body. Majority are linked to a core protein to form a proteoglycan
Chondroitin Sulfate
Most abundant GAG in the body Found in cartilage, tendons, ligaments and aorta Forms proteoglycan aggregates with hyaluronic acid.
Hyaluronate/Hyaluronic Acid
Different from other GAGs unsulfated not covalently attached to proteins Is a lubricant and shock absorber Lehninger Figure 7-24 Found in synovial fluid, vitreous humor, loose connective tissue
Additional GAGs
Keratin Sulfate present in cornea and connective tissue Dermatan Sulfate found in skin, blood vessels and heart valves Heparin
Proteoglycans
All GAGs, except for hyaluronate, are covalently attached to proteins to form proteoglycans. In cartilage, a core protein is linked to linear GAG chains. The negative charge of GAGs causes repulsion between molecules, generating a bottle brush appearance
Proteoglycans
Sugar Content >> Protein Content
Mucopolysaccharidoses
GAGs are normally degraded in lysosomes by specific enzymes Deficiency in any of these enzymes leads to accumulation of GAGs and a variety of symptoms such as skeletal and extracellular matrix deformities and mental retardation Hurler Syndrome -L-Iduronidase deficiency sx- corneal clouding mental retardation dwarfing Iduronate sulfatase deficiency X-linked inheritance sx- NO corneal clouding physical deformity mental retardation
Hunter Syndrome
Glycoproteins
Proteins modified by oligosaccharides Differ from proteoglycans carbohydrate chain is shorter, branched, fewer negative charges Function in cell surface recognition cell surface antigenicity (e.g. blood group antigens) extracellular matrix mucins of GI/GU tracts (biological lubricants)
Glycoproteins (contd)
O-glycosidic bonds carbohydrate is attached to hydroxyl group of serine or threonine N-glycosidic bonds carbohydrate is attached to the amide group of an asparagine side chain Occur as post-translational modifications the
Extended linear repetitions of disaccharides Central components of the extracellular matrix Hyaluronic Acid, Chondroitin Sulfate Glycoproteins Post-translational glycosylation of serine, threonine or asparagine residues Function in cellular recognition, cell surface antigenicity
Course Objective Review the biochemical processes most frequently tested on the NBDE
General Outline- 2nd hour Carbohydrate Metabolism Glycolysis Krebs Cycle Quiz
Digestion of Carbohydrates
Begins in the mouth - amylase breaks (14) bonds of starch and glycogen oligosaccharides In the small intestine - amylase produced by the pancreas further digests starch and glycogen oligo- and di-saccharidases made by intestinal mucosal cells catalyze the final digestion Small intestine absorbs
monosaccharides
Glycolysis
Anaerobic Cytosolic
Glycolysis
Anaerobic Cytosolic
Two phases Preparatory - 2 ATP per glucose Pay-off + 4 ATP per glucose Net 2 ATP per glucose
figure-14-2b
Glucose Glucose-6-Phosphate
ATP ADP
Regulation of Glycolysis
Phosphorylation of Glucose Phosphorylation of Fructose-6-Phosphate Generation of Pyruvate
Phosphorylation of Glucose commits glucose to inside cell two isoforms (Hexokinase vs Glucokinase) hormonal regulation
Hexokinase vs Glucokinase
Hexokinase- Ubiquitously expressed High affinity for glucose Low Km and Vmax Saturated at low [glucose] Glucokinase- Liver specific enzyme High Km and Vmax Saturated at high [glucose]
Regulation of Glycolysis
Phosphorylation of Glucose Phosphorylation of Fructose-6-Phosphate Generation of Pyruvate
Phosphofructokinase-1
Rate-Limiting Step Commits Glucose to Glycolysis Allosterically Regulated (-) ATP, citrate (+) AMP, F2,6BP
Fructose-6-Phosphate
ATP
ADP
Fructose-1,6-Bisphosphate
Hormonally Regulated
Regulation of Glycolysis
Phosphorylation of Glucose Phosphorylation of Fructose-6-Phosphate Generation of Pyruvate
Pyruvate Kinase
Feed-forward Regulation (+) F1,6,BP Large (-) G Essentially Irreversible Hormonally Regulated
Phosphoenolpyruvate
ADP
ATP
Pyruvate
Fates of Pyruvate
Anaerobic Cytosolic
Anaerobic Conditions
Pyruvate is converted to lactate In the absence of oxygen, glycolysis is the only means of generating ATP. Regenerates the NAD+ required by glyceraldehyde-3-phosphate dehydrogenase during the payoff phase of glycolysis Anaerobic bacteria also rely on this pathway to maintain ATP production
Aerobic Conditions
Anaerobic Conditions
TCA Cycle
Pyruvate Dehydrogenase
Converts pyruvate to CO2 and Acetyl CoA TCA cycle Multi-subunit enzyme requires several cofactors A Thiamine pyrophosphate Coenzyme NAD+ FAD Lipoic Acid
Energy is harnessed in the form of NADH Negative allosteric regulation by Acetyl CoA and NADH
(mitochondrial matrix)
-Ketoglutarate Dehydrogenase
Analogous to Pyruvate Dehydrogenase Multi-subunit enzyme requires several cofactorsA Thiamine pyrophosphate Coenzyme NAD+ FAD Lipoic Acid
TCA Cycle
Occurs in the mitochondrial matrix Citrate synthase is the rate-limiting step Sites of NADH synthesis Isocitrate dehydrogenase - Ketoglutarate dehydrogenase Malate dehydrogenase Site of FADH2 synthesis Succinate dehydrogenase (member of the ETC) Site of GTP synthesis Succinyl-CoA synthetase
TCA Cycle
For each Acetyl CoA 2 CO2 1GTP 3 NADH 1 FADH2
ETC
ATP Produced
by both substrate level phosphorylation and oxidative phosphorylation
Catabolism of Macromolecules
Converges on the formation of Acetyl CoA
Occurs exclusively in the mitochondrial matrix None of the reaction intermediates are consumed For each Acetyl CoA oxidized to 2 CO2 3 NADH 1 FADH2 1 GTP Reduced equivalents will be passed to the ETC for ATP synthesis Cycle is inhibited by high-energy substrates
Aerobic Conditions
Anaerobic Conditions
TCA Cycle
Pyruvate Dehydrogenase
Converts pyruvate to CO2 and Acetyl CoA TCA cycle Multi-subunit enzyme requires several cofactors A Thiamine pyrophosphate Coenzyme NAD+ FAD Lipoic Acid
Energy is harnessed in the form of NADH Negative allosteric regulation by Acetyl CoA and NADH
TCA Cycle
Occurs in the mitochondrial matrix Citrate synthase is the rate-limiting step Sites of NADH synthesis Isocitrate dehydrogenase - Ketoglutarate dehydrogenase Malate dehydrogenase Site of FADH2 synthesis Succinate dehydrogenase (member of the ETC) Site of GTP synthesis Succinyl-CoA synthetase
TCA Cycle
For each Acetyl CoA 2 CO2 1GTP 3 NADH 1 FADH2
ETC
http://www.dentistry.leeds.ac.uk/biochem/lecture/etran/etran.htm
Flow of Electrons Complex I Complex II Complex III Complex IV NADH dehydrogenase Succinate dehydrogenase Cytochrome bc1 complex Cytochrome oxidase (a+a3)
http://www.dentistry.leeds.ac.uk/biochem/lecture/etran/etran.htm
H+
H+
H+
e-
e-
e-
eH 2O
2H++ O2
NADH + H+ NAD+
NADH is oxidized by Complex I Electrons are passed down the chain H+ are pumped to the intermembrane space Only complexes I, III, and IV pump H+ Three H+ are pumped/NADH Oxygen is the terminal electron acceptor
H+
H+
e-
e-
eH 2O
http://www.dentistry.leeds.ac.uk/biochem/lecture/etran/etran.htm 2 2H++ O
FADH2+
FAD
FADH2 is oxidized by Complex II Complex II is Succinate Dehydrogenase Electrons are passed down the chain H+ are pumped to the intermembrane space Two H+ are pumped per FADH2
http://www.dentistry.leeds.ac.uk/biochem/lecture/etran/etran.htm
H+ ee-
H+ ee-
H+
H+
H+
ATPase
http://www.dentistry.leeds.ac.uk/biochem/lecture/etran/etran.htmO2 2H++
H 2O
NADH + H+
NAD+
Electron transfers generates a H+ gradient The energy generated by this gradient drives the synthesis of ATP
ADP + Pi
ATP
Insulin Signaling
Elevated blood glucose cells of the pancreas Insulin Insulin signaling increases expression of GLUT-4 receptors in insulin responsive tissues Skeletal muscle Glycogen Adipose tissue Triacylglycerol (TAG) Insulin promotes Glucose Uptake Glycolysis Glycogen Synthesis HMP shunt Triacylglycerol Synthesis Insulin signal Dephosphorylation of enzymes Enzymes activated by insulin are dephosphorylated in the active state
Structure of Insulin
Synthesized as Preproinsulin (RER) Loses its signal sequence to become Proinsulin Disulfide bonds link Chain A and Chain B (RER) Loses its C-peptide to become Insulin (Golgi)
Insulin Receptor
Cell-surface receptor Contains intrinsic tyrosine kinase activity Autophosphorylates Activates kinases and phosphatases Insulins effects are mediated by dephosphorylation
Lehninger Figure 12-6
Insulin Signaling
Increases Glucose uptake Glycogen Synthesis Protein Synthesis Fat Synthesis Decreases Gluconeogenesis Glycogenolysis Lipolysis Activates/Deactivates Enzymes
Alters Gene Expression Increases Glucose Transport
H 2O
Protein phosphatase
Insulin
Insulin
Activates/Deactivates Enzymes
Insulin
Activates/Deactivates Enzymes Alters Gene Expression
Insulin
Activates/Deactivates Enzymes Alters Gene Expression Increases Glucose Transport
Glucose uptake Glycogen Synthesis Protein Synthesis Fat Synthesis Gluconeogenesis Glycogenolysis Lipolysis
Alters enzyme activity Activates Glycogen Synthase Alters gene expression Upregulates genes of glycolysis Promotes GLUT-4 translocation skeletal muscle & adipose tissue
Glucagon Signaling
Low blood glucose cells of the pancreas Glucagon Promotes the mobilization of blood glucose glycogenolysis gluconeogenesis Glucagon promotes protein and/or fat catabolism Glucagon signal phosphorylation of enzymes Enzymes activated by glucagon are phosphorylated in the active state Actions of Glucagon can also be performed by Epinephrine
Glucagon/Epinephrine Signaling
Activates G protein Activates Adenylate Cyclase cAMP Activates cAMP dependent protein kinase, Protein Kinase A
()
Glucagon/Epinephrine Signaling
Leads to the phosphorylation of enzymes Phosphorylation activates glycogen phosphorylase a Glycogenolysis
Glucagon
cAMP
Glucagon/Epinephrine Signaling
Protein Kinase A
ATP ADP
H 2O
Protein phosphatase
Insulin
Glucagon
cAMP Protein Kinase A
ATP ADP
Glycogen is Degraded
Glycogen P Phosphorylase a (active)
ADP ATP H 2O Pi Insulin Protein phosphatase
Protein phosphatase
Insulin
Glycogen Synthesis
Skeletal muscle and liver take up glucose and store it as glycogen [(14) and (16)] Glycogen synthesis occurs during the well-fed state Insulin favors glycogen synthesis Glucagon opposes glycogen synthesis
Glycogen Synthesis
glucose 6- phosphate glucose 1- phosphate
phosphoglucomutase
UDP- glucose is the substrate for glycogen synthesis Anomeric carbon of sugar is activated by attachment to UDP
+ +
Glycogen Synthesis
Two enzymes glycogen synthase glycogen-branching enzyme
Glucagon
cAMP
Glucagon Signals through the activation of Protein Kinase A Glycogen Synthase is inactive in the phosphorylated form
Protein Kinase A
ATP ADP
H 2O
Protein phosphatase
Insulin Signals through the activation of Protein Phosphatase Glycogen Synthase is active in the dephosphorylated form
Insulin
NADPH
6-Phosphogluconate
6-Phosphogluconate Dehydrogenase NADP+
NADPH
Products of the Pentose Phosphate Pathway NADPH Fatty Acid and Cholesterol Synthesis
Counter Free Radicals RBCs are exposed to oxygen generated free radicals Glutathione functions to eliminate these free radicals NADPH is required to keep glutathione in the reduced state G6PD deficiency free radical damage hemolytic anemia
Lehninger Box-14-03
Occurs in cells not needing the pentose sugars Functions to regenerate glucose 6-phosphate
Glucose 6-Phosphate Glucose 6-Phosphate can be used in Glycolysis Glycogen Synthesis HMP Shunt
General Outline- 2nd hour Carbohydrate Metabolism Glycogen Synthesis Hexose Monophosphate Shunt Glucose Mobilization Glycogenolysis Gluconeogenesis
Glucagon:Insulin Ratio
Activates two processes to maintain blood glucose Glycogenolysis skeletal muscle liver Gluconeogenesis liver kidney
Glycogenolysis
Glycogenolysis functions during the first 24 hours of fasting Glucose from liver glycogen blood glucose Glucose from muscle glycogen energy for the myocyte
Glycogenolysis
Glucagon Activates
Insulin Inactivates
Glycogenolysis
Insulin
Glucagon
cAMP Protein Kinase A
ATP ADP
Glycogen is Degraded
Glycogen P Phosphorylase a (active)
ADP ATP H 2O Pi Insulin Protein phosphatase
Protein phosphatase
Insulin
Glycogenolysis
Glucose 1-phosphate>>Glucose
Activated by Glucagon
Glycogen is mobilized within four hours of eating Stimulated by Glucagon phosphorylates and activates glycogen phosphorylase kinase glycogen phosphorylase phosphorylates and inactivates glycogen synthase Occurs in liver blood glucose and muscle energy
Glucagon:Insulin Ratio
Activates two processes to maintain blood glucose Glycogenolysis skeletal muscle liver
Gluconeogenesis
The de novo synthesis of glucose Occurs in the liver>>kidney during fasting Substrates include all intermediates of glycolysis all intermediates of the TCA cycle lactate glycerol -ketoacids of glucogenic amino acids Is NOT a reversal of Glycolysis Must bypass Hexokinase Phosphofructokinase Pyruvate Kinase
To counter the irreversible steps of glycolysis Four enzymatic steps Four ATP and Two GTP
Enzymes in Glycolysis
Hexokinase
Enzymes in Gluconeogenesis
Glucose-6-Phosphatase Fructose 1,6-bisphophatase Phosphoenolpyruvate Carboxykinase Pyruvate Carboxylase
Phosphofructokinase
Pyruvate Kinase
Mitochondrion
Pyruvate + CO2
ATP
Acetyl CoA
ADP + Pi
Oxaloacetate
Malate
Cytosol
Malate
Oxaloacetate
GTP PEP Carboxykinase GDP
Phosphoenolpyruvate + CO2
PEP
Reverse Glycolysis
Fructose 6-Phosphate
Bypasses the irreversible phosphofructokinase-1 reaction Stimulated by High ATP Low AMP Inhibited by Elevated AMP Fructose 2,6-bisphosphate
Fructose 6 - phosphate
Bypasses the irreversible hexokinase reaction Only occurs in the LIVER and KIDNEY Glucose 6-phosphatase is essential for glucose to be released into the blood stream Without this enzyme, glucose generated by glycogenolysis and gluconeogenesis can not be released to the body
Substrates for Gluconeogenesis All intermediates of glycolysis All intermediates of the TCA cycle Lactate Glycerol -Ketoacids of glucogenic amino acids
When glucose is limiting Muscles go into anaerobic metabolism Lactate Fats are catabolized Glycerol and fatty acids Proteins are catabolized amino acids
Cori Cycle
Active muscle mobilizes glycogen stores glucose Glucose is metabolized by glycolysis under anaerobic conditions lactate Lactate enters blood liver Gluconeogenesis resynthesizes glucose from lactate Glucose enters blood muscle Lehninger Figure 23-18 Gluconeogenesis occurs in LIVER and KIDNEY Glucose can be metabolized or stored as muscle glycogen
Regulation of Gluconeogenesis
Glucagon Phosphorylates enzymes deactivation Pyruvate Kinase PFK-2 Induces the expression of PEP carboxykinase Substrate Availability Glucogenic amino acids increase rate Reciprocal Control + Acetyl CoA Activates pyruvate carboxylase Inhibits pyruvate dehydrogenase Inhibits Fructose 1,6-bisphosphatase - AMP Activates PFK-1
General Outline- 2nd hour Lipid Structure and Function Cholesterol and Fatty Acids Compartmental Barrier Derivatives of Cholesterol and FAs Fatty Acid Synthesis
Cholesterol
Abundant in cell membrane Also functions as a precursor Bile acids Vitamin D Steroid hormones Transported in Lipoprotein Particles Most abundant in LDL>HDL>VLDL Found in the diet and synthesized in all tissues Especially liver, adrenal cortex and reproductive tissues
Lehninger Figure-10-16
Amphipathic Hydrophobic AND Hydrophilic Saturated Unsaturated Bonds No Double Bonds One Double
>90% are esterified Cholesteryl esters Phospholipids Triacylglycerides (TAG) Saturated Unsaturated Free FAs circulate bound to ALBUMIN ( during fasting)
Function Oxidized for energy by most tissues (esp. liver and muscle) Structural roles (phospholipids & glycolipids) Precursor of prostaglandin Major Energy Store (TAG)
Unsaturated Saturated
Essential
Triacylglycerol (TAG)
May also be referred to as triglycerides Major energy store found in adipocytes Three fatty acids esterified to glycerol Neutral Fat When mobilized FA Acetyl CoA TCA Glycerol glycolysis or gluconeogenesis
Lehninger Figure-10-2
Lehninger Figure-17-1
Chylomicrons
Largest, least dense lipoprotein Predominantly dietary triacylglycerol Generated in enterocytes lymph blood stream tissues Tissues will consume or store the dietary triacylglycerol Remnants return to the liver release their cholesterol degraded in lysosomes
Lehninger Figure-17-2
Lipoprotein Particles
Chylomicrons Transport TAG from the diet to the tissues Apo B-48 LDL Particles Transport cholesterol to the tissues Receptor-mediated endocytosis Apo B-100 VLDL Particles Transport TAG synthesized by the liver to the tissues HDL Particles Transport cholesterol to the liver for elimination
Lehninger Figure-10-6
Glycerophospholipids
General Structure Glycerol backbone Two Fatty Acids Phosphate Group + Polar Head Group
Sphingolipids
General Structure Sphingosine backbone One Fatty Acid Head Group Sphingomyelin Sphingolipid prevalent in myelin Also contains choline Deficiency in degradation Sphingolipidoses
Lehninger Figure-10-12
Sphingolipidoses
Tay-Sachs Hexosaminidase A deficiency GM2 gangliosides accumulate Neurodegeneration Cherry-red Macula Gauchers Glucocerebrosidase deficiency Glucocerbrosides accumulate Hepatosplenomegaly Osteoporosis of long bones Niemann-Pick disease Sphingomyelinase deficiency Spingomyelin accumulation Hepatosplenomegaly Neurodegenerative
Facilitated diffusion occurs down gradient does not require energy e.g. glucose uptake Active transport against gradient requires energy e.g. Na/K ATPase
Three forms of transport simple diffusion facilitated diffusion require transporter active transport
down gradient
Steroid Hormones
Derived from cholesterol Nonpolar, lipophilic Circulate bound to a carrier protein Enter cells by simple diffusion Bind to INTRAcellular receptors Ultimately reach the nucleus to alter gene expression
Lehninger Figure-10-19
Eicosanoids
Prostaglandins, Thromboxanes, Leukotrienes
Have Hormone-Like Properties Physiological and Pathological Responses Vasoconstriction/Vasodilation Contraction/Relaxation of smooth muscle Bronchoconstriction and Anaphylaxis (Leukotrienes) Differences From Hormones Act locally Not stored Short half-life Produced by all tissues in small amounts Derived from arachidonic acid, a fatty acid component of the cell membrane
Lehninger Figure-10-18
Arachidonic Acid liberated from phospholipid by Phospholipase A2 Converted by Cyclooxygenase to Prostaglandins and/or Thromboxanes Converted by Lipooxygenase to Leukotrienes
A large portion of fatty acids come from the diet chylomicrons tissues Fatty acids can also be synthesized from carbohydrates and proteins Primarily occurs in the liver and mammary glands, and, to a lesser extent, in adipose tissue Requires Acetyl CoA, ATP, Biotin, and NADPH Fatty Acid Synthesis Occurs in the Well-Fed State High Insulin:Glucagon Ratio
Mitochondrion
Oxaloacetate + Acetyl CoA
Citrate Synthase
CoA
Citrate
Cytosol
CoA, ATP
Citrate
ATP-Citrate Lyase
ADP + Pi
LCFA-CoA
ADP + Pi
Citrate
Malonyl CoA
Glucagon
cAMP
Protein Kinase A
ATP ADP
H 2O
Protein phosphatase
Insulin
Epinephrine
cAMP
Protein Kinase A
ATP ADP
Protein phosphatase
Insulin
Triacylglycerol
HormoneSensitive Lipase
DHAP
Mitochondria
-Oxidation
Acetyl CoA
Brain and RBCs can NOT use FAs
TCA
Lehninger Figure-17-6
ATP
Lehninger Figure-17-7
ETC
Lehninger Figure-17-12
Glucagon and Insulin also regulate Hormone-Sensitive Lipase Active when phosphorylated by Glucagon Inactive when dephosphorylated by Insulin
Synthesis
Greatest flux through pathway Hormonal State favoring pathway Major tissue site Subcellular location Carriers of acyl groups Oxidation/reduction cofactors Two-carbon donor/ product Activator Inhibitor Product of Pathway After carbohydrate rich meal High Insulin:Glucagon Liver Cytosol Citrate NADPH Malonyl CoA (donates acyl) Citrate LCFA-CoA Palmitate
Degradation
In starvation Low Insulin:Glucagon Muscle, Liver Mitochondria Carnitine NAD+, FAD Acetyl CoA (product) Malonyl CoA Acetyl CoA
Formed during fasting and in Diabetes Type I Production in excess of rate of use Ketonemia Ketonuria
Ketoacidosis
Brain can utilize Ketone Bodies RBCs can NOT use Ketone bodies
Protein Metabolism
Protein catabolism occurs in three scenarios after a protein-rich meal during normal turnover of proteins during prolonged fasting or starvation Proteins catabolism results in the generation of free amino acids. Amino acids are degraded to carbon skeletons substrate for glucose ammonia toxic byproduct that is eliminated
Lehninger Figure-18-3
Denaturation of Proteins
Digestive Proteases
Stomach Parietal Cells HCl, Intrinsic Factor Chief Cells Pepsinogen Pepsin Pancreas Trypsinogen Chymotrypsinogen Procarboxypeptidases Intestine Enteropeptidase (Enterokinase) Cleaves trypsinogen to active trypsin Trypsin cleaves and activates itself and all other zymogens
Digestive Proteases
Trypsin (Serine Protease) Cleaves C-terminal to basic amino acids Chymotrypsin Cleaves C-terminal to aromatic amino acids Carboxypeptidases Remove the C-terminal residue of a peptide Aminopeptidases Remove the N-terminal residue of a peptide
Amino acids are absorbed from the small intestine and enter the portal circulation The portal vein feeds the liver where amino acids are further catabolized Catabolism of amino acids requires two enzymes Transaminase (aminotransferase) Glutamate Dehydrogenase Amino acids are degraded to carbon skeletons (-ketoacids) ammonia (toxic byproduct)
Aminotransferase
Catalyzes the first step in amino acid catabolism Transfers amino group from amino acids to -ketoglutarate Glutamate and the -keto-acid of the amino acid degraded are formed Requires pyridoxal phosphate as a cofactor (from Vitamin B6) Nearly all amino acids funnel their amino groups to glutamate
Lehninger Figure-18-4
Glutamate Dehydrogenase
Catalyzes the second step in amino acid catabolism Only enzyme that can use either NAD or NADP Generates NH4+ and -Ketoglutarate
Lehninger Figure-18-7
Glutamate
NH4+ + -Ketoglutarate
-Ketoglutarate
NADH (NADPH)
-ketoacids
-NH2 of glutamate
NAD+ (NADP+)
Alanine Aminotransferase
Alanine -Ketoglutarate Alanine is deaminated to pyruvate Pyruvate is the -ketoacid of alanine
Pyruvate
Glutamate
Aspartate Aminotransferase
Oxaloacetate Glutamate Oxaloacetate is the -ketoacid of aspartate Formation of aspartate is favored during degradation Aspartate -Ketoglutarate Aspartate is required for the urea cycle
Lehninger Figure18-10
The detoxification of ammonia occurs in the liver Glutamine is the predominant carrier of amino groups from most tissues Alanine transports amino groups from skeletal muscle and delivers its -ketoacid, pyruvate, to the liver. Glutamate NH3 and Aspartate
Urea Cycle
Lehninger Figure18-10
Occurs in both the mitochondria and cytosol Functions to convert ammonia to a non-toxic form Urea The two amino groups of urea are acquired from free ammonia (NH3) and aspartate The rate-limiting step Carbamoyl phosphate synthetase I
Lehninger Figure18-10
Oxaloacetate from the TCA cycle is the alpha-ketoacid of aspartate Fumarate is generated in the urea cycle and returns to the mitochondrial matrix for the TCA
Amino acids are processed transamination oxidative deamination Amino groups are collected in the form of free ammonia and aspartate Free ammonia and aspartate donate their amino groups to the formation of non-toxic urea
Amino Acid
Degradation of Phenylalanine
Lehninger Figure-18-23
Phenylketonuria (PKU)
Inability to hydroxylate phenylalanine to tyrosine Primarily caused by deficiency of phenylalanine hydroxylase Phenylalanine is degraded by an alternate pathway phenylketones Phenylketones in urine musty odor Symptoms MR, failure to walk or talk, seizures, hyperactivity Neonatal Diagnosis
Lehninger Figure-18-25
General Outline- 2nd hour Synthesis of Non-Essential Amino Acids Amino Acids as Substrates Water-Soluble Vitamins Fat Soluble Vitamins
Aminotransferase
-ketoacids
-NH2 of glutamate
NAD+ (NADP+)
Transamination and oxidative deamination are readily reversible, according to the cells needs.
Histamine GABA Inhibitory Neurotransmitter Deficiency leads to seizures Potent Vasodilator Released from mast cells during allergic response Production in the stomach stimulates acid production
VITAMINS
Water-soluble
Fat-soluble
Vitamin A Vitamin D Vitamin E Vitamin K
NonB-complex
Ascorbic acid (Vitamin C)
B-complex
Energy-releasing
Thiamine (B1) Riboflavin (B2) Niacin (B3) Biotin Pantothenic acid
Hematopoeitic
Folic Acid Vitamin B12
Other
Pyridoxine (B6) Pyridoxal Pyridoxamine
Biotin
Coenzyme in carboxylation reactions Required for pyruvate carboxylase acetyl CoA carboxylase Biotin Deficiency is Rare Pantothenic Acid Component of coenzyme A Required for TCA, fatty acid synthesis and degradation Pantothenate Deficiency is Poorly Characterized
Folate
Found in leafy green vegetables Functions in one-carbon transfers as coenzyme tetrahydrofolate Important in the synthesis of amino acids, purines and thymine
Example Oxaloacetate + Glutamate Aspartate + -ketoglutarate Serine Pyruvate + NH3 Histidine Histamine + CO2
Vitamin
Folic Acid Vitamin B12 Vitamin C Vitamin B6 Vitamin B1 Niacin Riboflavin Biotin Pantothenic Acid
Function
Transfer of 1-carbon units Synthesis of purines and thymine Degradation of oddnumbered fatty acids Antioxidant Cofactor for Hydroxylations Cofactor for a.a. catabolism Cofactor for oxidative decarboxylation reactions Electron Transfer Electron Transfer Carboxylation reactions Acyl carrier
Deficiency
Megaloblastic Anemia Neural Tube Defects Megaloblastic Anemia Neuropsychiatric Sx Scurvy (loose teeth & Poor wound healing) Rare Beriberi Wernicke-Korsakoff Pellagra (3 Ds) Rare Rare Rare
Vitamin A (Retinol)
Retinoids class of compounds related to Vitamin A required for vision, growth and epithelial tissues Deficiency causes night blindness and xerophthalmia Excess Vitamin A leads to toxicity teratogenic
Vitamin E
Primarily functions as an antioxidant, preventing the nonenzymatic oxidation of compounds by molecular oxygen or free radicals Deficiency in adults is rare Least toxic of the fat-soluble vitamins (ADEK)
Vitamin D
Two precursors Ergocalciferol (Acquired through the diet) 7-Dehydrocholesterol (Synthesized in the skin) Precursors are activated by hydroxylation position 25- in the liver position 1- in the kidney Active 1,25 dihydroxycholecalciferol Synthesis of active Vitamin D is stimulated by parathyroid hormone in response to low blood calcium.
Lehninger Figure-10-20
Vitamin D (contd)
Vitamin D functions to maintain blood calcium levels increases calcium absorption in the intestine limiting loss of calcium through the kidney promoting bone resorption in conjunction with PTH Vitamin D Deficiency Rickets (in children) Collagen matrix of bone is formed but is not completely mineralized Soft, pliable bones Osteomalacia (in adults) Existing bones are demineralized and are much more susceptible to fracture Vitamin D deficiency can be a result of insufficient UV exposure dietary deficiency advanced renal disease
Vitamin K
Vitamin K is essential for post-translational modification of clotting factors II, VII, IX, and X Blood clotting factors are synthesized in the liver where they undergo Vitamin K-dependent carboxylation of their glutamate residues. Glutamate carboxylation is required for clotting factors to interact with activated platelets at the site of a wound. Modified glutamate has an overall charge of -2. Warfarin is a competitive inhibitor used as an anticoagulant in patients at risk of heart attack or stroke. Deficiency of Vitamin K is rare in adults microbes in gut synthesize Vitamin K
X
VII
Cells expose Tissue Factor
XIIa IXa
+
XII
XIa
VIIa
IX
VIIIa
Prothrombin
Xa + Va
Factors II, VII, IX and X are Vitamin K dependent Factors V and VIII are coenzymes
Function
Reproduction Vision Maintenance of epithelial tissue Calcium Uptake Antioxidant Cofactor for -Carboxylation of glutamate in clotting factors
Deficiency
Impotence Night Blindness Xerophthalmia Rickets (children) Osteomalacia (adults) Rare RareCompetitively inhibited by Warfarin