Glycolysis and Gluconeogenesis

Learning Goals:
1. Learn the reactions of glycolysis and gluconeogenesis, understand the purposes served by each.
2. Understand regulation of glycolysis and gluconeogenesis
3. Understand differences between skeletal muscle and liver in metabolic role and control mechanisms.

I. The BIG Picture

a. Glycolysis- metabolizes glucose to pyruvate/lactate with modest energy yield; can run at high speed
without oxygen
b. Gluconeogenesis replenishes glucose consumed by glycolysis
i. Liver: gluconeogenesis maintains the blood glucose level
ii. Muscle: gluconeogenesis replenishes muscle glycogen consumed in earlier bursts of muscle
activity.
c. Control: irreversible steps, effected by metabolites/hormones
II. Overview of glycolysis and gluconeogenesis
a. Glycolysis
i. Universal metabolic pathway
ii. Six carbon glucose to two three carbon pyruvates
iii. In presence of oxygen: end product pyruvate metabolized to CO2 by the TCA Cycle
iv. Glucose = ideal substrate for rapid energy production because of solubility/ability of glycolysis to
metabolize glucose without oxygen
b. Gluconeogenesis
i. Synthesizes glucose from smaller precursors
ii. Important in liver- responsible for maintaining a constant blood glucose concentration
iii. Muscle does NOT carry out significant gluconeogenesis
III. The Glycolytic Pathway: Glycolysis is stimulated by insulin and inhibited by glucagon
a. Stage 1
i. Energy expended: 2 ATP  phosphorylate the original glucose molecule
ii. Diphospho sugar cleaved to yield three carbon phospho compounds
b. Stage 2
i. Energy yielding segment: 2 ATP generated per three carbon intermediate = 4 molecules of ATP =
net gain of 2 ATP
c. Glycolysis contains 2 irreversible steps: PFK-1 and pyruvate kinase
i. PFK-1: catalyzes conversion of fructose-6-phosphate to fructose-1,6-bisphosphate, subject to
allosteric regulation [ATP], [AMP], [ADP], [citrate]
ii. Important allosteric activator of PFK-1: fructose-2,6-bisphosphate (NOT a metabolic
intermediate, only regulatory molecule); sets overall rate of PFK-1  glycolysis
1. Formed from fructose-6-phosphate by phosphofructokinase-2
2. Broken down to fructose-6-phosphate by fructose bisphosphatase-2
d. Fructose-1,6-bisphosphate cleaved to yield dihydroxyacetone phosphate and glyceraldehyde-3-phosphate
by the action of aldolase (reversal of aldol concensation, requires active site lysine  Schiff base)
e. DHAP converted to GAP by triosephosphate isomerase  both proceed through second stage of
glycolysis as DHAP.
f. Conversion of GAP to 1,3-bisphosphoglycerate by glyceraldehyde-3-phosphate dehydrogenase
(oxidation-reduction reaction): creates high energy phosphate bond, NADH
i. Enzyme inactivated by heavy metals that coordinate to cysteine SH group
g. Phosphoglycerate kinase: hydrolysis of mixed acid anhydride of 1,3-bisphosphoglycerate to formation of
ATP  substrate level phosphorylation; makes 3-phosphoglycerate
i. RBCs: 1,3-bisphosphoglycerate also utilized to generation 2,3-DPG by action of
bisphosphoglycerate mutase
h. Phosphoglycerate mutase moves phosphate
i. Enolate dehydrates 2-phosphoglycerate to yield phosphoenol pyruvate (phosphoenol bond = high energy)
j. Pyruvate kinase catalyzes transfer of phosphate from PEP to ADP  pyruvate and ATP (substrate level
phosphorylation)
i. Pyruvate kinase activated by fructose-1,6-bisphosphate
ii. Pyruvate kinase inhibited by high ATP and alanine
 iii. Pyruvate kinase covalently regulated by phosphorylation (cAMP-dependent protein kinase)
1. Pyruvate kinase is ACTIVE when unphosphorylated
2. Pyruvated kinase is INACTIVE when phosphorylated
k. Alternate fates of pyruvate depending on O2 availability (requirement to regenerate NAD from NADH)
i. Anaerobic glycolysis: NAD regenerated by production of lactate
ii. Aerobic glycolysis NAD regenerated by shuttling reducing equivalents into the mitochondrion for
subsequent oxidative phosphorylation
1. Glycerol phosphate shuttle
l. Fructose can enter glycolysis: Tissues other than liver- fructose directly phosphorylated to fructose-6-P by
hexokinase, Liver- fructose enters glycolysis through the fructose-1-P pathway
m. Galactose enters glycolysis as glucose-6-P by way of the galactose-glucose interconversion pathway.
n. Defects in pyruvate kinase  hemolytic anemia arising from compromised ability to maintain adequate
cellular ATP in RBCs
IV. The Gluconeogenic Pathway
a. Pyruvate converted to OAA in the mitochondrion by pyruvate carboxylase  malate/asp shuttle  OAA
decarboxylated at the expense of GTP by PEP carboxykinase  PEP
b. PEP metabolized backwards through the glycolytic pathway to fructose-1,6-bisphosphate
c. Fructose-1,6-bisP converted to fructose-6-P and Pi by fructose-1,6-bisphosphatase (gluconeogenic
specific enzyme) **crucial regulatory site**
d. Glucose-6-P converted to glucose and Pi by glucose-6-phosphatase
e. Cori Cycle: muscle  liver