CHAP 17: GLYCOLYSIS & THE OXIDATION OF PYRUVATE by ZKPJ

BIOMEDICAL IMPORTANCE GLYCOLYSIS CAN FUNCTION UNDER  Hexokinase is allosterically INHIBITED by its
 Brain - in prolonged fasting, it gets 20% of its ANAEROBIC CONDITIONS product, G-6-P
energy from Ketone Bodies  Present in most tissues
 Muscle contracts in anaerobic medium:
 Glycolysis - major pathway for glucose  Low Km, Low Vmax
 Glycogen disappears
metabolism ------------------------------------------------------------
 Lactate appears
 Occurs in the cytosols of ALL cells Muscle contracts in aerobic medium:  Glucokinase

 Can function either:  Lactate does not appear  An isoenzyme of hexokinase, which is found
 Aerobically or in liver cells, iselt pancreatic cells
 Pyruvate is the major end product of
 Anaerobically glycolysis  High Km, High Vmax
(depending on the availability of:  Function is to remove glucose from the
 Pyruvate is further oxidized to CO2 & H2O
Oxygen & the E.T.C.) hepatic portal blood following a meal
 When Oxygen supply is short:
 Erythrocytes  Regulates the [glucose] available to
 Mitochondrial reoxidation of NADH formed
 lack mitochondria, peripheral tissues
during glycolysis is impaired
 are completely dependent on glucose  Stimulated by Glucose, Insulin
 NADH is reoxidized by reducing pyruvate to
as their metabolic fuel  Provides more G-6-P that is required for
lactate, so permitting glycolysis to continue
 Metabolize glucose anaerobic glycolysis
 Anaeorobic Glycolysis is LIMITING:
glycolysis  G-6-P is used for glycogen synthesis and
 Limits the amount of ATP formed per mole of
 To oxidize Glucose: lipogenesis
glucose oxidized
 Oxygen  More glucose is required to be metabolized to  Also found in Beta-cells of pancreas, where it
 Mitochondrial Enzyme Systems: functions to detect high conc. of glucose
meet energy demand
i. Pyruvate dehydrogenase More glucose phosphorylated by Glucokinase,
REACTIONS OF GLYCOLYSIS Increased Glycolysis,
complex I. ENERGY INVESTMENT PHASE
ii. Citric Acid Cycle Increased formation of ATP
 Glycolysis from glucose to lactate:  Increased [ATP]i leads to closure of ATP-K+
iii. Respiratory Chain
 Glycolysis is the principal route for carbohydrate channel exchange
metabolism.  ALL enzymes of Glycolysis are CYTOSOLIC  Results to membrane depolarization
 Can provide ATP for Skeletal muscles when 1. Glucose enters Glycolysis by Phosphorylation to  Opening of voltage-gated Calcium
Oxygen supply is insufficient Glucose-6-Phosphate (G-6-P), channel
 Cardiac muscles are adapted for aerobic  catalyzed by Hexokinase  Influx of Ca2+ leads to fusion of insulin
performance secretory granules with the cell
 Low glycolytic activity membrane
 Poor survival under conditions of  Release of Insulin
ischemia  G-6-P is an important compound at the junction of
Glycolysis > Pyruvate > Lactate > Gluconeogenesis several metabolic pathways:
 Lactic Acidosis - results from various causes,  Glycolysis
including impaired activity of pyruvate  Gluconeogenesis
dehydrogenase, especially in thiamin (Vitamin  Pentose Phosphate Pathway
B1) deficiency  Glycogenesis
 Glycogenolysis
 Phosphorylation of Glucose to G-6-P is ------------------------------------------------------------
IRREVERSIBLE 2. G-6-P is isomerized to Fructose-6-phosphate
CHAP 17: GLYCOLYSIS & THE OXIDATION OF PYRUVATE by ZKPJ
 catalyzed by Phosphohexose isomerase  DHAP (Liver and Adipose tissue)
3. F-6-P is Phosphorylated to fructose  Used for triglyceride synthesis
1,6-biphosphate  Deficiency of glycolytic enzymes lead to
 catalyzed by Phosphofructokinase Hemolytic Anemia
a) Phosphofructokinase is IRREVERSIBLE II. ENERGY GENERATION PHASE
under physiological conditions 5. Glycolysis continues with the oxidation of G-3-P to
b) RATE LIMITING STEP 1,3-bisphophate 8. 2-Phosphoglycerate is converted to
c) Inhibited by high levels of ATP, citrate  catalyzed by G-3-P dehydrogenase phosphoenolpyruvate
d) Activated by high levels of AMP, fructose a) G-3-P dehydrogenase is NAD dependent  Catalyzed by Enolase
2,6 biphosphate - most potent b) 1st oxidation-reduction reaction of a) Dehydration (removal of water)
glycolysis b) REVERSIBLE
c) NADH formed must be reoxidized by: c) Enolase is inhibited by Fluoride
i. NADH-linked coversion of pyruvate to d) Enolase is dependent on the presence of
lactate either Mg2+ or Mn2+ ions
ii. Oxidation of NADH via the respiratory
chain

4. Fructose 1,6-bisphosphate is cleaved by

Aldolase into two triose phosphates:
i. Glyceraldehype-3-phosphate (G-3-P)
ii. Dihyrdoxyacetone phosphate
the two can be interconverted
phosphotriose isomerase
 Aldolase A
 Readily REVERSIBLE
 Not regulated
 Aldolase B (Liver and Kidney)
 Cleaves Fructose 1,6-Bisphosphate
 Metabolism of fructose
9. Phosphoenolpyruvate is converted to Pyruvate
by  Catalyzed by Pyruvate kinase
a) Phosphate group of Phosphoenolpyruvate is
transferred to ADP > 1 ATP
b) Pyruvate kinase is IRREVERSIBLE under
physiological conditions
c) SECOND substrate-level phosphorylation
 The availability of Oxygen determines which of the
d) Arsenic Poisoning: two pathways is followed:
i. Arsenate competes with Pi as substrate  Anaerobic conditions:
for G-3-P dehydrogenase in this reaction  NADH cannot be reoxidized through the
ii. Give rise to 1-arseno-3-phosphoglycerate ETC
WITHOUT forming ATP  Pyruvate is reduced to lactate cataylzed
6. Phosphate group is transferred from by lactate dehydrogenase
1,3-bisphosphoglycerate to ADP > 1 ATP  This permites oxidation of NADH,
 Catalyzed by Phosphoglycerate kinase permitting another molecule of glucose
 FIRST substrate-level phosphorylation to undergo glycolysis
7. 3-phosphoglycerate is ISOMERIZED to
 Aerobic conditons:
2-phosphoglycerate  Pyruvate is transported into
 Catalyzed by phosphoglycerate mutase mitochondria
CHAP 17: GLYCOLYSIS & THE OXIDATION OF PYRUVATE by ZKPJ
 Pyurvate undergoes oxidative TISSUES THAT FUNCTION UNDER HYPOXIC  LACTATE CONSUMPTION:
decarboylation to acetyl-CoA CONDITIONS PRODUCE LACTATE  Heart & Liver: Lower NADH/ NAD+ ratio than
 Then oxidation to CO2 in the CAC  Glycolysis in RBCs ALWAYS terminates in lactate, during exercise
 The reducing equivalents from the because the subsequent pyruvate oxidation are  Liver: Pyruvate is converted to glucose or
NADH formed in glycoylsis are taken mitochondrial, and RBCs LACK mitochondria oxidized in the Citric Acid Cycle cycle
up into mitcohondira for oxidation via  Tissues that derive their energy from glycolysis  Heart: exclusively oxidizes Lactate to CO2
 Malate-aspartate shuttle or and produce lactate include: and H2O via the Citric Acid Cylce
 Glycerophosphate shuttle  Brain  LACTIC ACIDOSIS
 Pyruvate Kinase Deficiency  Gastrointestinal tract  Occur with collapse of the circulatory system
 85% of patients with genetic defects of  Renal medulla 1. Failure to bring adequate amounts of
glycolytic enzymes  Retina oxygen to tissues
 2nd most common cause of enzymatic  Skin 2. IMPAIRED oxidative phosphorylation
related hemolytic anemia  The Liver, Kidneys, and Heart normally 3. Decreased ATP synthesis
 Restricted to RBCs, producing mild to take up Lactate and oxidize it, but  Solution: use of the Anaerobic system
severe hemolytic anemia produce it in under HYPOXIC  Oxygen debt: excess oxygen required to
 Severity: depends on the degree of enzyme CONDITIONS recover from a period when the availability of
deficiency  Increased Lactate production oxygen has been inadequate
 Covalent Modulation of Pyruvate Kinase Ex. vigorous exercise, septic shock, cancer cachexia GLYCOLYSIS IS REGULATED IN AT THREE STEPS
 Phosphorylation of cAMP-dependent INVOLVING NONEQUILIBRIUM REACTIONS
protein kinase leads to inactivation of Most is used in the Liver for Gluconeogenesis  3 Reactions are markedly EXERGONIC and
protein kinase in the Liver physiologically IRREVERSIBLE
i. LOW blood [glucose] => Glucagon Increase in metabolic rate to provide the ATP &  Hexokinase
secretion GTP needed  Phosphofructokinase
ii. Increased Glucagon levels  Pyruvate kinase
=> increase [cAMP]i  Accumulation of Lactate in Muscle causes the these 3 are the MAJOR sites of regulation of
iii. increase [cAMP]i => phosphorylation intracellular pH to drop => Muscle Cramps Glycolysis
& activation of Pyruvate Kinase  QUESTION: Potangina unsay may naa aning  Fructose
iv. PEP unable to continue glycolysis, Lactate oi??? (What’s so unique about Lactate?)  Enters glycolysis by phosphorylation to
enters Gluconeogenesis  ANSWER: F-1-Phosphate
v. Dephosphorylation of pyruvate  Lactate is a MAJOR precursor molecule  Bypasses the 3 major regulatory steps
kinase by phosphoprotein that can be used to form GLUCOSE in  Results to the formation of MORE
phosphatase => enzyme reactivation GLUCONEOGENESIS.  Pyruvate
Hormonal Regulation of Glycolysis  Skeletal muscle cells produce lactic  Acetyl-CoA
INSULIN GLUCAGON acid during exercise when the rate of than is required for ATP formation
Glucokinase + - glycolysis EXCEEDS the rate of  In the liver and adipose tissue, this leads to
Phosphofructokinase + - OXIDATIVE PHOSPHORYLATION INCREASE LIPOGENESIS
Pyruvate kinase + -  However, as mentioned above, RBCs  High intake in fructose = Obesity
ALWAYS produce lactic acid because
they lack the mitochondria required to
regenerate the NAD+ needed for
glycolysis
CHAP 17: GLYCOLYSIS & THE OXIDATION OF PYRUVATE by ZKPJ
I. IN ERYTHROCYTES, THE FIRST SITE OF ATP
FORMATION IN GLYCOLYSIS MAY BE BYPASSED
a) In Erythrocytes:
i. Reaction catalyzed by
phosphoglycerate kinase may be by
passed to some extent by the reaction THE OXIDATION OF PYRUVATE TO ACETLY-CoA IS THE
of Bisphosphoglycerate mutase
ii. This reaction catalyzes the formation
of 1,3-bisphosphoglycerate to
2,3-bisphosphoglycerate (2,3-BPG)
iii. 2,3-BPG gets hydrolyzed by
2,3-bisphosphoglycerate
phosphatase to 3-phosphoglycerate
+ Pi
b) This pathway involves NO NET YIELD OF ATP I. PYRUVATE DEHYDROGENASE IS REGULATED BY
from glycolysis END-PRODUCT INHIBITION & COVALENT
c) Binds to hemoglobin, decreasing its affinity MODIFICATION
for oxygen, so making oxygen more readily a) Pyruvate dehydrogenase
available to tissues i. Inhibited by its products:
1. Acetyl-CoA
IRREVERSIBLE ROUTE FROM GLYCOLYSIS TO THE CAC 2. NADH
 Pyruvate is transported into the mitochondrion by ii. Also regulated by phosphorylation of its
a proton symporter 3 serine residues on the Pyruvate
 Inside, it is OXIDATIVELY DECARBOXYLATED to dehydrogenase component of the
Acetyl-CoA by a multienzyme complex called multienzyme complex
Pyruvate Dehydrogenase Complex  Results to DECREASED activity
iii. Dephosphorylation
 Results to INCREASED activity
iv. Along with glycolysis, it is also inhibited
by adequate ATP available
v. Inhibited when fatty acids are being
oxidized
II. CLINICAL ASPECTS
a) ARSENIC POISONING
i. Toxicity: inhibition pyruvate
dehydrogenase
ii. React with the –SH groups (also with
mercuric ions)
iii. Results to accumulation of pyruvate
b) THIAMINE DEFICIENCY
i. Inhibit Pyruvate dehydrogenase
ii. Accumulation of Pyruvate
iii. Alcoholics: Pyruvic and Lactic Acidosis
c) Cofactors and Coenzymes used: (T.L.C.F.N.)
i. Thiamine pyrophosphate (TPP) From
Thiamine (vitamin B12)
ii. Lipoic Acid
iii. Coenzyme A from Pantothenate
iv. FAD(H2) from Riboflavin
v. NAD(H) from Niacine
(read na lang sa book hahaha)
CHAP 17: GLYCOLYSIS & THE OXIDATION OF PYRUVATE by ZKPJ
III. SUMMARY: ENERGY YIELD FROM GLYCOLYSIS
a) ANAEROBIC:
i. 1 Glucose = 2 Lactate = 2 ATP
ii. No net production / consumption of
NADH
b) AEROBIC:
i. 1 Glucose = 2 Pyruvate = 2 ATP (net
gain)
ii. 1 Glucose = 2 NADH