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Lecture 5-3: Regulation of Glycolysis

Glycolysis is a central pathway of glucose metabolism and therefore of


carbohydrate metabolism. Its regulation is central to energy management.

• Introduction
• Basic mechanisms – intersections of pathways
• Physiological questions
• Major sites of regulation
• Regulation in liver
• Regulation in cardiac muscle
• Alcohol metabolism
• Pathways
• Clinical correlations
• Clinical scenario: Chronic alcohol consumption

All images are from “Marks et al., 2nd Edition, Copyright © 2005 Lippincott &
Williams, A Wolters Kluwer Co., All rights reserved” unless otherwise noted.

Glycolysis is central to both


catabolic and anabolic metabolism

(storage) Note: G6P is not transported back


across the plasma membrane
Glucose-6-P

• Catabolic reactions:
•Source of ATP
• Anabolic reactions:
Complex Glycolysis •Pyruvate is precursor for
carbohydrates fatty acid biosynthesis
NADPH
and alanine
ATP •Pentose phosphates
precursor of nucleotides.

TCA Cycle
Figure 2 of the Section Five Introduction, Mark’s et al.
Major pathways of glucose metabolism

(storage)

Glucose-6-P

Complex Glycolysis
carbohydrates NADPH
ATP

TCA Cycle
Figure 2 of the Section Five Introduction, Mark’s et al.

Glucose

Anaerobic
Glycolysis
Lactate
Alternate fates of pyruvate

Figure 22.5: glycolysis occurs


exclusively in the cytosol

Glucose + 2 ADP + 2 Pi →
2 Lacate + 2 ATP + 2 H2O + 2 H+

The overall ATP yield for the complete oxidation


of one mole of glucose to CO2 is 36 – 38 moles
Fig. 22.9
x2
x2
A brief note about ‘shuttles’

Fig. 22.8: Malate - aspartate shuttle Fig. 22.7: Glycerol 3-phosphate shuttle

Review of Unit 4…

Regulation of enzymatic activity – A pathway is


only as fast as its slowest step
• Enzymes may be activated (+) or inhibited (-)
• The concentration of enzymes may be increased or decreased
through the regulation of transcription or translation
¾ Induction (+) or repression (-)
• Mechanisms of regulation include
¾ Allosteric
¾ Hormonal
Physiological roles
• Tissue specific responses in glucose homeostasis
¾ Liver is a “maintainer organ” – its principal role is to maintain
homeostasis
¾ Muscle is a “consumer organ” – its principal role is to convert
chemical to mechanical energy
Pi
Glucose 6-
phosphatase
Synthesis of Glucokinase
is induced by insulin

The major sites of


glycolysis regulation
in the liver

↑ Insulin : glucagon ratio –


positively regulates PFK-1 and PK

Figure 36.1: Regulation of glucokinase,


PFK-1, and pyruvate kinase in the liver.

Product inhibition

The major sites of


glycolysis regulation
in the skeletal muscle

Figure 22.12: Regulation of hexokinase


and PFK-1 in skeletal muscle.
Fig. 31.14 -
Differences in Km
between the liver
enzyme glucokinase
and hexokinase, the
enzyme that is found
in other tissues and
catalyzes the same
reaction

Regulatory considerations with regard to glucokinase


• Km (S0.5) ≈ 7 - 10 mM - physiological blood glucose is
5 - 10 mM (80-160 mg/dl)
• Glucokinase enzyme induced by increased ratio of insulin : glucagon
• Balance of glucokinase and glucose 6-phosphatase activities
(G 6-P → glucose + Pi)

Pi
Glucose 6-
phosphatase

The major sites of


glycolysis regulation
in the liver

↑ Insulin : glucagon ratio –


positively regulates PFK-1 and PK

Figure 36.1: Regulation of glucokinase,


PFK-1, and pyruvate kinase in the liver.
Regulation of Phosphofructokinase-1 (PFK-1)
PFK-1 catalyzes Fructose 6-Phosphate → Fructose 1,6 Bisphosphate

• Allosteric regulation
of PFK-1 by AMP to
a lower Km (higher
affinity) +AMP

Fig. 22.14 Regulation of PFK-1 by AMP,


ATP and fructose 2,6-bisphosphate.

Regulation of Rest
glycolysis in ~ 20%
Exercise
cardiac muscle
by AMP

adenylate
kinase
2 ADP ←⎯⎯→ AMP + ATP
~ 300%

Marks Figure 22.13: Changes in


ATP, ADP and AMP concentrations
in skeletal muscle during exercise.
Regulation of Phosphofructokinase-1 (PFK-1)

• Allosteric regulation of
PFK-1 by F2,6-BP to
a lower Km (higher
affinity)
• F2,6-BP is more
effective activator of
PFK-1, mole for mole,
than AMP

Fig. 22.14 Regulation of PFK-1 by AMP,


ATP and fructose 2,6-bisphosphate.

Fructose 2,6-bisphosphate (F2,6-BP) regulates PFK-1


• Fuctose 2,6-BP is synthesized by Phosphofructokinase-2 (PFK-2)
• The level of F 2,6-BP is controlled by phosphorylation of PFK-2

• F2,6-BP is not an glycolysis


intermediate
• F2,6-BP regulates PFK-1 in
both liver and adipose tissue
• [F2,6-BP] increases when
the insulin : glucagon ratio
increases since glycolysis in
these tissues is the carbon
source for the synthesis of
triacylglycerols
Fructose 2,6-Bisphosphate is synthesized and
degraded by a single enzyme, PFK-2

PFK-2
has both
of these
activities

Phosphofructokinase-2 (PFK-2) is a bifunctional enzyme. The ratio of


activities is changed by phosphorylation and dephosphorylation of PFK-2.

Coordinate regulation of liver PFK-2


• The two activities of PFK-2 are controlled by Protein
Kinase A and phosphoprotein phosphatase

↑ cAMP

Higher PFK-2
Higher PFK-2
phosphatase
kinase activity
activity
results in results in less
more F 2,6-BP F 2,6-BP
Activation of glycolysis by Fructose 2,6-Bisphosphate

• In liver and adipose tissue


after a high carbohydrate
meal (↑ Insulin : Glucagon)
• PFK-2 is dephosphorylated P
phosphorylase
by protein phosphorylase PFK-2 PFK-2
• The kinase activity of PFK-2
increases
• Therefore, levels of F2,6-BP F-2,6-BP F-6-P
increase
• F2,6-BP allosterically
activates PFK-1 PFK-1
• Glycolysis is stimulated F-6-P F-1,6-P
allowing glucose to be
converted to triacylglycerols
for storage

Inhibition of glycolysis in the LIVER by glucagon


or epinephrine via a signal transduction cascade
↓ Insulin : Glucagon

From Figure 28.8:


The steps of a signaling cascade…
activation of protein kinase A (PKA) →
phosphorylation of phosphofructokinase-2 (PFK-2) →
increase in the fructose 2,6-bisphosphatase activity →
less fructose 2,6-bisphosphate (F 2,6-BP) →
phosphofructokinase-1 (PFK-1) activity decreases →
…resulting in less glycolysis in the liver
The response of PFK-2 to
hormones is tissue specific
For example, the hormone epinephrine mobilizes fuels
during acute stress -
• Liver: A ‘maintainer’ organ, provides plasma glucose
by limiting glycolysis: Epinephrine → elevated cAMP,
phosphorylates bifunctional enzyme → increases
PFK-2 phosphatase activity → less F 2,6 BP
• Cardiac muscle: A ‘consumer’ organ, increases
glycolysis to produce pyruvate for energy production:
Epinephrine → elevated cAMP, phosphorylates
bifunctional enzyme → increases PFK-2 kinase
activity → more F 2,6 BP

Tissue specific enzyme isozymes

Liver
PFK-2
Liver phosphoryation favors phosphatase activity

Cardiac
Muscle
PFK-2 Muscle phosphoryation favors kinase activity

Regulation of liver pyruvate kinase by phosphorylation status. P = a serine residue


phosphorylated by protein kinase A. (Alternative splicing yields different proteins.)
Liver:
Pyruvate kinase

Liver:
Pyruvate kinase
Alcohol (Ethanol) is a fuel

Ethanol is principally metabolized


in the liver providing as many as 13
moles of ATP per mole of ethanol

Alcohol (Ethanol) is a fuel

Use of ethanol as a motor fuel on a


large scale raises environmental and
food supply concerns
Alcohol metabolism
Alcohol dehydrogenase
(Cytosolic enzyme)

(Toxic)

Fig. 25.1

Aldehyde dehydrogenase
(Mitochondrial enzyme)

(Nontoxic)
Fig. 25.2

Alcohol metabolism

TCA cycle
Fatty Acid Synthesis
Clinical scenario: Metabolic effects of
chronic ethanol consumption
• Summary of metabolic pathways
• Ethanol is both lipid and water soluble and thus is
readily absorbed by passive diffusion
• Ethanol is principally metabolized in the liver with as
many as 13 ATP synthesized per molecule
• Products are acetaldehyde, acetate and acetyl CoA
• Acetyl CoA →TCA cycle or fatty acid synthesis in the liver
• Acetaldehyde is toxic
• Reducing equivalent NADH and H+ produced
• Nutritional Effects
• Chronic alcohol consumption results in decreased
adsorption of vitamins B1 (thiamin) and B12
• Behavioral Effects – (Acetaldehyde)

Nutritional effects?

?
Source: Carlsberg brewery website
• The enzymes of alcohol metabolism are
a family of isozymes
• Variation in the isozyme proportions affects
• rate of alcohol clearance,
• degree of inebriation,
• side effects of alcohol consumption and
• susceptibility to alcohol-induced liver disease
• Points 2 – 4 are directly related to the
concentration of the acetaldehyde
intermediate

Genetic variation of aldehyde


dehydrogenase (ALDH)
• The common allele of ALDH a low Km resulting in
a high affinity for its substrate acetaldehyde
• Since even low concentrations of acetaldehyde
(toxic) are converted to acetate (nontoxic), the
concentration of the intermediate rarely exceeds
20 μM even during intoxication.
• An ALDH isozyme present with ~ 40% frequency
in East Asian populations has a high Km resulting
in low affinity for its substrate acetaldehyde
• High acetaldehyde concentrations result from
minimal alcohol consumption
• Accumulation of toxic acetaldehyde results in
facial flushing, nausea and vomiting
• Possession of this isozyme is associated with
protection against alcoholism
Genetic variation of alcohol
dehydrogenase (ADH)
• Isozymes of alcohol dehydrogenase that have a
low Km result in metabolic changes similar to an
acetaldehyde dehydrogenase isozyme with high
Km and thus low affinity.
• The concentration of a reaction intermediate is the
result of its influx and efflux…

Why isn’t ethanol a ‘good’ fuel?

The products of its metabolism


alter the flow of metabolites…
Alcohol metabolism favors
ketosis and triglyceride
synthesis through the
excessive production of NADH