Metabolism

GLYCOGENESIS
• Pathway by which glucose ultimately is converted into glycogen.
• Important in hepatocytes  liver major site synthesis and storage
• 7%  wet weight of the liver
• Liver glycogen carbohydrates reenter blood stream
• Maintains blood glucose homeostasis
• Other sitesskeletal muscle and, adipose tissue (lesser extent)
• Skeletal muscle <1% wet wt of tissues
• Body’s glycogen  75% Stored in muscles (make greater portion of body than
liver)
• Glycogen stores in muscle energy sourcephysical exertion.
• Glucose phosphorylated entering the cell phosphate ester at the 6-carbon of the
glucose (glucose 6 phosphate)
• Catalyzing Enzyme (muscle cells)  phosphate transfer from ATP hexokinase
(allosteric enzyme)
• Glucose phosphorylation (liver) (glucokinase or hexokinase D)
• Hexokinase  negatively modulated by glucose 6-phosphate, but glucokinase is not.
 GLYCOGENESIS
• Glucokinase  higher Km than hexokinase
• Kmglucose to its phosphate form at a higher velocity
• liver is not dependent upon insulin for glucose transport
into the cell
• Hexokinase  not induced by insulin
• Glucokinase is inducible by insulin
• Glucokinase activity is below normal values in type 1
diabetes patients.
• low glucokinase activity  liver cell’s inability to rapidly take
up and metabolize glucose
• The hexokinase/glucokinase reactionenergy
consuming .
 GLYCOGENESIS
• Glycogenesis initiated presence of glucose 6-
phosphate.
• The phosphorylation of glucosekeeps the level of
free glucose lowenhances the entry of glucose
 GLYCOGENESIS
• The phosphate is transferred from the 6-carbon of the glucose to
the 1-carbon in a reaction catalyzed by the enzyme
phosphoglucomutase.
• In the next reaction, energy derived from the hydrolysis of the α-β-
phosphate anhydride bond of uridine triphosphate (UTP to UMP)
• uridine monophosphate coupled to the glucose 1-phosphate to
form uridine diphosphate glucose (UDP-glucose). UDP-glucose
pyrophosphorylase
• Glucose is incorporated into glycogen as UDP-glucose catalyse by
glycogen synthase requires some preformed glycogen as a primer
• Initial glycogen is formed by binding a glucose residue
to a tyrosine residue of a protein called glycogenin
 GLYCOGENESIS
• Additional glucose residues are attached by glycogen
synthase to form chains of up to eight units.
• In muscle the protein stays in the core of the glycogen
molecule.
• liver more glycogen molecules than glycogenin
molecules the glycogen must break off of the protein
• Glycogen synthase exists in an active (dephosphorylated)
form and a less active (phosphorylated) form.
• Insulin facilitates glycogen synthesis by stimulating the
dephosphorylation of glycogen synthase.
 GLYCOGENESIS
• When six or seven glucose molecules are added to the
glycogen chain the branching enzyme transfers them to a C(6)
—OH group
• Glycogen synthase cannot form the α 1-6 bonds of the branch
points.
• Branching within the glycogen important increases the
molecule’s solubility and compactness.
• Branching makes available many nonreducing ends glucose
residues can be cleaved rapidlyenergy glycogenolysis
 GLYCOGENOLYSIS
• The breakdown of glycogen into individual glucose units, in the form of glucose
1-phosphate, is called glycogenolysis.
• Regulated by hormones glucagon (pancreatic origin) acts in liver and
adipose tissues
catecholamine hormone epinephrine (produced in
the adrenal medulla) liver and muscles
• Both hormones function through the second messenger cAMP

• Glucagon and epinephrine function antagonistically to insulin in regulating the

balance between glucose and glycogen.
• energy is needed glucose units glycogen phosphorolysis

• Glycosidic bonds cleaved by phosphate addition.

• Products  glucose 1-phosphate and the remainder of the intact glycogen chain
• Catalyzed by  glycogen phosphorylase phosphorylase a (a
phosphorylated active form)
phosphorylase b (a dephosphorylated, inactive form)
 GLYCOGENOLYSIS
• Glycogen phosphorylase (phosphorylated by) Phosphorylase b
kinase.
• Dephosphorylated by phosphorylase a phosphatase
• Rate of glycogen breakdowndepends relative activity of
phosphorylase a and phosphorylase b.
• Breakdown of liver and muscle glycogen Complex process
covalent regulation (phosphorylationdephosphorylation regulation)
allosteric regulation by modulators (enzymes and
hormones)
 GLYCOGENOLYSIS
• Covalent regulation
 Strongly influenced by the hormones glucagon
and epinephrine
• Glucagon acts in the liver and adipose tissue
• Epinephrine  acts in the liver and muscle
• Both these hormones stimulating phosphorylase b
kinase promoting formation of the more active
(“a”) form of the enzyme.
Phosphorylase b kinase  mediated through
cAMP (cellular concentration increased by
same enzymes)
 GLYCOGENOLYSIS
• Allosteric regulation of phosphorylase involves the
positive modulator AMP
• ATP competes with AMP for the allosteric site of the
enzyme High ATP levels  keep it in its inactive form.
• Induces conformational change in the inactive “b fully
active “b” form
• No covalent (phosphorylation) regulation is involved in
allosteric modulations.
Glycogen phosphorylase cleaves α 1-4 glycosidic bonds
not α 1-6 bonds.
• Debranching enzyme cleaves the α 1-6 bond at the
branch point.
 GLYCOGENOLYSIS
• Glucose 1-phosphate glucose 6-phosphate
through glucose phosphate isomerase
• Glucose 6-phosphatecan enter glycolysis or free
glucose (liver kidney)
• Glucose 6-phosphate  free glucose require
glucose 6-phosphatase (only in liver and kidney
cells)
• Thus, the liver (but not muscle) can control the
concentration of glucose in the blood
 GLYCOLYSIS
• Glucose two units of pyruvate, a triose
• Metabolism of glucose Aerobic or Anaerobic
(depending upon available oxygen)
• Anaerobic conditions Oxygen debt pyruvate 
lactate (strenuous exercise)
• Lactate muscle bloodstreamliverglucose.
• Anaerobic glycolysissole source of energy for
erythrocytes (no mitochondria)
• Brain and GI tract produce energy through glycolysis.
 GLYCOLYSIS
• Under aerobic conditions pyruvate transported to
mitochondria participate in TCA cycleoxidized to
CO2 and H2O (demands an ample supply of oxygen)
• Glycolytic enzymes function within the cytoplasmic
matrix of the cell.
• Enzymes catalyzing the TCA cycle in mitochondria
• Pyruvate must enter the mitochondrion for
complete oxidation.
 GLYCOLYSIS
• Glycolysis provids initial sequence of reactions
(to pyruvate)  TCA cyclelarge quantities of ATP.
• Step1 The hexokinase/glucokinase reaction
consumes 1 mol. ATP/mol glucose. Hexokinase in
muscle (but not glucokinase) is negatively regulated
by the product of the reaction glucose 6-phosphate.
Glucokinase in liver (but not hexokinase) is induced
by insulin.
 GLYCOLYSIS
• Step 2 Glucose phosphate isomerase (also called
hexose phosphate isomerase) catalyzes this
interconversion of isomers—glucose 6-P to fructose
6-P.
• Step 3 The phosphofructokinase reaction, an
important regulatory site, is modulated (by
allosteric mechanisms) negatively by ATP and citrate
and positively by AMP and ADP.
• Also regulated hormonally by glucagon (induction)
 GLYCOLYSIS
• Step 4 Aldolasesplitting a hexose (fructose)
bisphosphate into two triose phosphates,
glyceraldehyde 3-phosphate and dihydroxyacetone
phosphate
• (DHAP). The prefix “bis” means two phosphates are
present, on different carbons atoms.
• The prefix “di” means that the two phosphates are
attached to each other and to a single carbon atom.
 GLYCOLYSIS
• Step 5 Glyceraldehyde 3-phosphate and
dihydroxyacetone phosphate interconvertable
triosephosphate isomerase.
• In an isolated system, the equilibrium favors DHAP
formation.
• In the cellular environment, however, it is shifted
completely toward producing glyceraldehyde 3-
phosphate removed from the equilibrium by the
subsequent reaction catalyzed by glyceraldehyde 3-
phosphate dehydrogenase.
 GLYCOLYSIS
• Step 6 Glyceraldehyde 3-phosphate oxidizedcarboxylic
acid, while inorganic phosphate is incorporated as a high-
energy anhydride bond.
• The enzyme is glyceraldehyde 3-phosphate dehydrogenase,
which uses NAD+ as its hydrogen-accepting cosubstrate.
Under aerobic conditions, the NADH formed is reoxidized to
NAD+ by O2 through the electron transport chain
in the mitochondria.
The reason why O2 is not necessary to sustain the reaction
of converting glyceraldehyde 3-P to bis-P-glycerate is
that under anaerobic conditions the NAD+ consumed is
restored by a subsequent reaction converting pyruvate to
lactate.
 GLYCOLYSIS
• Step 7 This reaction, catalyzed by phosphoglycerate
kinase, exemplifies a substrate-level
phosphorylation of ADP.
• Two ATPs are synthesized because glucose (Hexose)
makes two trioses.
• Step 8 Phosphoglyceromutase catalyzes the transfer
of the phosphate group from the number 3 carbon
to the number 2 carbon of the glyceric acid.
• Step 9 Dehydration of 2-phosphoglycerate enolase
introduces a double bond imparts high energy to
the phosphate bond.
 GLYCOLYSIS
• Step 10 Phosphoenolpyruvate (PEP) donates its
phosphate group to ADPcatalyzed by pyruvate kinase.
• This is the second site of substrate-level
phosphorylation.
• Step 11 The lactate dehydrogenasetransfers two
hydrogens from NADH and H+ to pyruvatereducing it
to lactate.
• NAD+ is formed in the reaction and can replace the
NAD+ consumed earlier under anaerobic conditions.
 GLYCOLYSIS
• Under normal, aerobic conditions, pyruvate enters
the mitochondrion for complete oxidation.
• 3rd option pyruvate  alanine (transamination)
a reaction by which pyruvate acquires an amino
group from the amino acid glutamate
GLYCOLYSIS
GLYCOLYSIS
 GLYCOLYSIS
• Step 12
• Fructose is directly phosphorylated
by hexokinase D (glucokinase) to form fructose 6-
phosphate.
• unimportant reaction
• The hexokinase reaction is slow and occurs only in
the presence of high levels of fructose.
• Fructose to fructose 1-P is the major means by
which fructose is converted to glycolysis
metabolites.
GLYCOLYSIS
 GLYCOLYSIS
• Step 14 Galactose to galactose 1-P through enzyme
galactokinase
• Step 15 Galactose 1-P to Glucose 1-P through
galactose 1-phosphate uridyl transferase.
• Epimerase, UDP-galactose UDP-glucose glucose
1-phosphate (previous reaction)
• OR incorporated into glycogen by glycogen synthase
(glycogenesis)
• Can also enter the glycolytic pathway as glucose 6-
phosphate (Glucose phosphate Isomerase)
• Free Glucose (glucose 6-Phosphatase) only liver.
GLYCOLYSIS
 TCA cycle
• tricarboxylic acid cycle (TCA cycle)=Krebs cycle =citric acid cycle
• final catabolic pathway  produces 90% energy along with CO2
and H2O
• TCA cycle intermediates gluconeogenesis amino acids by
transamination
• located within the mitochondrial matrix
• oxidation reactions=dehydrogenations removal of two
hydrogens (½H2) to an acceptor cosubstrate such as NAD+ or
FAD
• NADH and FADH2 readily reoxidized by O2
through the electron transport chain, located in the
mitochondrial inner membrane
 TCA cycle
• TCA cycle produces most of the carbon dioxide
through decarboxylation
•
Kreb Cycle
 Kreb Cycle
• pyruvate dehydrogenase complex multienzyme
system and various cofactors, with the enzymes and
cofactors
• Cofactorscoenzyme A (CoA), thiamin pyrophosphate
(TPP), Mg2+, NAD+, FAD, and lipoic acid.
• Four vitamins required for activity of complex
pantothenic acid (B5) (a component of CoA), thiamin
(B1), niacin (B3), and riboflavin (B2).
• Pyruvate dehydrogenase complex  pyruvate
decarboxylase, dihydrolipoyl dehydrogenase, and
dihydrolipoyl transacetylase.
 Kreb Cycle
• Net effect of the complex decarboxylation and
dehydrogenation of pyruvate
• Reoxidation of NADH by electron transportapprox
3 mol of ATP
• Reaction allosterically negatively modulated acetyl
CoA and by NADH
positively by ADP and Ca2+.
 Condensation of acetyl CoA with oxaloacetate initiates
the TCA cycle reactions.
 Kreb Cycle
• Net effect of the complex decarboxylation and
dehydrogenation of pyruvate
• Reoxidation of NADH by electron transportapprox
3 mol of ATP
• Reaction allosterically negatively modulated acetyl
CoA and by NADH
positively by ADP and Ca2+.
 Condensation of acetyl CoA with oxaloacetate initiates
the TCA cycle reactions.
 Kreb Cycle
1) The formation of citrate from oxaloacetate and
acetyl CoA is catalyzed by citrate synthase. The
reaction is regulated negatively by ATP.
2) The isomerization of citrate to isocitrate cis
aconitate as an intermediate. The isomerization,
catalyzed by aconitase, involves dehydration
followed by sterically reversed hydration, resulting
in the repositioning of the —OH group onto an
adjacent carbon.
 Kreb Cycle
3 Catalyzed by  isocitrate dehydrogenase
 First of four dehydrogenation reactions
 Energy supplied  reoxidation NADH.
 First loss of CO2by spontaneous decarboxylation
of oxalosuccinate (reaction intermediate)
 Positively modulated by ADP and negatively
modulated by ATP and NADH.
 Kreb Cycle
4 Decarboxylation and dehydrogenation of α-
ketoglutarate mechanistically identical to the
pyruvate dehydrogenase
 α-ketoglutarate dehydrogenase reaction, NAD+ serves
as hydrogen acceptor, and a second carbon is lost as
CO 2.
 The pyruvate dehydrogenase, isocitrate
dehydrogenase, and α-ketoglutarate dehydrogenase
reactions account for the loss of the three carbons
from pyruvate as CO2.
 Kreb Cycle
5 Energy conserved in the thioester bond of succinyl
CoA.  hydrolysis  by succinyl thiokinase releases
sufficient energy to drive the phosphorylation of
guanosine diphosphate (GDP) by inorganic phosphate.
 GTPserve as phosphate donorfor example
gluconeogenesis or glycogenesis. GTP can transfer its
γ-phosphate to ADP to form ATP.
 Kreb Cycle
6 The succinate dehydrogenase reaction uses FAD
instead of NAD+ as hydrogen acceptor. The FADH2
is reoxidized by electron transport to O2, but only
about two ATPs are formed by oxidative
phosphorylation instead of three.
7 Fumarase incorporates H2O across double bond
of fumarate to form malate.
 Kreb Cycle
8 The conversion of malate to oxaloacetate
completes the cycle.
NAD+  acts as hydrogen acceptor (malate
dehydrogenase).
This reaction is the fourth site of reduced
cosubstrate formation (3-NADH and 1-FADH2) and
thus results in additional energy release in the
cycle.
 ATPs Produced By Complete Glucose Oxidation
complete oxidation of glucose glycolytic and TCA cycle pathways.
C6H12O6 + 6 O2 6 CO2 + 6 H2O + energy
 By convention 1NADH= 3 ATPS
1 FADH2=2 ATPs
 Actual number of ATPs formed from NADH is closer
to 2.5; for FADH2, it is 1.5.
 under aerobic conditions
1) glucose two pyruvates,
produces four ATPs by substrate-level phosphorylation.
However, two ATPs are used in the pathway, producing a
net of two ATPs. (2 ATPs)
 ATPs Produced By Complete Glucose Oxidation
2) The two NADHs glyceraldehyde 3-phosphate
dehydrogenase reaction
yield 4 or 6 ATPs 6 for malate-aspartate
shuttle system
4 for glycerol-3-phosphate shuttle system
 Acetyl CoA Oxidation and Tricarboxylic Acid Cycle Intermediates
TCAsteady supply of four-carbon units needed  oxidize all of the acetyl
CoA produced to CO2 and H2O
In absence of four-carbon intermediates, ketoacidosis results
 Acetyl CoA is produced by fatty acid oxidation and amino acid catabolism, as
well as from the pyruvate derived from glycolysis leads to an imbalance
between the amounts of acetyl CoA and oxaloacetate
 oxaloacetate and/or other TCA cycle intermediates that can form
oxaloacetate must be replenished in the cycle.
 Oxaloacetate, fumarate, succinyl CoA, and α-ketoglutarate can all be formed
from certain amino acids
 conversion of pyruvate to oxaloacetate is called an anaplerotic (filling-up)
process because of its role in restoring oxaloacetate to the cycle.
 Acetyl CoA Oxidation and Tricarboxylic Acid Cycle Intermediates
absence of four-carbon intermediates,
ketoacidosis results.
Acetyl CoA is produced fatty acid oxidation and
amino acid catabolism, as well as from the pyruvate
derived from glycolysis
increase in acetyl CoA leads to an imbalance
between the amounts of acetyl CoA and oxaloacetate
single most important mechanism for ensuring an
ample supply of oxaloacetate  catalyzed by
pyruvate carboxylase
 Acetyl CoA Oxidation and Tricarboxylic Acid Cycle Intermediates
The conversion of pyruvate to oxaloacetate is called
an anaplerotic (filling-up) process.
 Acetyl CoA Oxidation and Tricarboxylic Acid Cycle Intermediates
The conversion of pyruvate to oxaloacetate is called
an anaplerotic (filling-up) process.
 NADH in Anaerobic and Aerobic Glycolysis:
The Shuttle Systems
shuttle systems are specific to certain tissues.
more active malateaspartate shuttle functions in the
liver, kidney, and heart
glycerol 3-phosphate shuttle functions in the
brain and skeletal muscle.
 Glycerol -Phosphate Shuttle System
shuttle systems are specific to certain tissues.
more active malateaspartate shuttle functions in the
liver, kidney, and heart
glycerol 3-phosphate shuttle functions in the
brain and skeletal muscle.
Glycerophosphate produced by glycolysis is oxidized
by two different glycerophosphate dehydrogenases,
one in the cytoplasm and the other on the outer
face of the inner mitochondrial membrane
 Glycerol -Phosphate Shuttle System
intramitochondrial reoxidation of glycerol 3-
phosphate is catalyzed by glycerol phosphate
dehydrogenase, which uses FAD instead of NAD+
as hydrogen acceptor.
 if the glycerol 3-phosphate shuttle is in effect, only
four ATPs are formed aerobically per mole of
glucose by oxidative phosphorylation
 Malate-Aspartate Shuttle System
freely permeable to the inner mitochondrial
membrane.

GLYCOLYSIS
GLYCOLYSIS
GLYCOLYSIS
GLYCOLYSIS
 ETC
• Complex 1
• Contains NADH dehydrogenase
(46) several polypeptide chains
1FMN
 NADH-CoQoxidreductase
coenzyme Q
Several Fe-S clusters with additional
iron molecules
iron molecules bind with the sulfur-containing amino acid
cysteine.
iron transfers one electron at a time cycling between Fe+2/Fe+3
 ETC
• Net result transfer of electrons and hydrogen from
NADH to form reduced coenzyme Q
 transfer of hydrogen ions (4) from
the matrix side of the inner
mitochondrial membrane to the
cytosolic side of the inner
membrane.
 ETC
• Complex II
• integral part of the inner mitochondrial membrane.
 Succinate dehydrogenase
 FAD protein
Fe-S clusters
FADH 2  oxidized with one electron transfers through
the Fe-S centers to reduce coenzyme Q to coenzyme QH2
oxidation of FADH2 through the electron
transport chain results in the formation of approximately
2 molecules of ATP
 ETC
• Complex III (Cytochrome C Oxidoreductase )
• coenzyme Q passes its electrons to cytochrome
c in the third complex of the electron transport
chain in a pathway known as the Q cycle.
• Three cytrochromes
Cytochrome C1 (Contains one heme group)
Cytochrome B (contains two heme group)
Rieske center ( 2Fe-2S group)
 ETC
• Complex III (Cytochrome C Oxidoreductase )
• coenzyme Q passes its electrons to cytochrome
c in the third complex of the electron transport
chain in a pathway known as the Q cycle.
• Three cytrochromes
Cytochrome C1 (Contains one heme group)
Cytochrome B (contains two heme group)
Rieske center ( 2Fe-2S group)
 Fat Metabolism
• Mitochondrial Transfer of Acyl CoA
 FATE OF PROTEN
• First step in the metabolism of amino acids not
used for the synthesis of proteins or nitrogen- containing compounds
is the removal of the amino group (transaminiation/deamination)
• Some amino acids that are more commonly deaminated include
glutamate,
histidine, serine, glycine, and threonine
• Deamination reactions are generally lyases, dehydratases, or
dehydrogenases
• Ammonia is readily used by periportal hepatocytes for urea synthesis.
• Transamination involve the transfer of an amino group from one
amino acid to an amino acid
carbon skeleton or α-keto acid (an amino acid without
an amino group)
 FATE OF PROTEN
• First step in the metabolism of amino acids not
used for the synthesis of proteins or nitrogen-
containing compounds is the removal of the amino
group (transaminiation/deamination)
• Deamination reactions are generally lyases,
dehydratases, or dehydrogenases
 FATE OF PROTEN
• Transamination reactions are catalyzed by enzymes
called aminotransferases (PLP dependent) (tyrosine
minotransferase, branched-chain minotransferases,
alanine aminotransferase, and aspartate
aminotransferase.
• (ALT and AST) most active of the
aminotransferases and involve three key amino
acids: alanine, glutamate, and aspartate.
• Aminotransferases are found in varying
concentrations in different tissues.
 FATE OF PROTEN
• AST found mainly in heart
• ALT found mainly in liver
• AST and ALT as well as alkaline phosphatase and
lactate dehydrogenase high plasma concentration
 indicate liver damage
• Heart attack High plasma concentration AST and
a specific MB form of creatine kinase (also called
creatine phosphokinase)
 FATE OF PROTEN
• ALT transfers amino groups from alanine to
an α-keto acid (e.g., α-ketoglutarate), forming
pyruvate and another amino acid (e.g., glutamate)
• AST transfers amino groups from aspartate also to an
α-keto acid (e.g., α-ketoglutarate), yielding
oxaloacetate and another amino acid (e.g., glutamate)
• Reactions are reversible. Because glutamate and
α-ketoglutarate readily transfer and/or accept amino
groups, these compounds play central roles in amino
acid metabolism.
 FATE OF PROTEN
• lysine, histidine, and threonine  do not take part in
transamination