GLUCONEOGENESIS
Definition: Biosynthesis of glucose
from simpler molecules, primarily
pyruvate and its precursors.
Why do we produce glucose?
• Need to maintain glucose levels in a narrow
range in blood.
• Some tissue-- brain,RBC,lens,adrenal medulla
and muscles in exertion use continuous glucose
supply.
•Hepatic glycogen depleted in 10 – 18 hrs of
fasting, subsequently gluconeogenesis is the
only source.
Where is glucose synthesized?
• Liver(90%) & kidney(10%)- major sites
• Requires both cytosolic & mitochondrial
enzymes.
• Pyruvate, lactate, glycerol & - ketoacids from
glucogenic aa are the main precursors.
What are the substrates?
Pyruvate - major precursor
Lactate– from muscle
glycerol from certain lipids.
some amino acid carbon skeletons- from diet or
breakdown of muscle protein during starvation-
most important is alanine
TCA cycle intermediates
propionate from breakdown of fatty acids and
amino acids.
Cost: The production of glucose is
energy expensive.
Input: 2 pyruvate + 4 ATP + 2 GTP +
2 NADH
Output: glucose + 4 ADP + 2 GDP +
2 NAD++ 6 Pi
Energy barriers obstruct a simple reversal of
glycolysis.
Three Glycolysis reactions have such a large
negative DG that they are essentially irreversible.
- Hexokinase (or Glucokinase)
- Phosphofructokinase
- Pyruvate Kinase.
These steps must be bypassed in Gluconeogenesis.
SUBSTRATES FOR GLUCONEOGENESIS
Pyruvate
-major precursor.
Lactate - the primary source for pyruvate.
NAD+ NADH+H+
.
Lactate Pyruvate
Lactate Dehydrogenase
CORI’S CYCLE
-- In muscle, lactate is produced in great
quantities during exertion
-- This excess lactate cannot be further oxidized
in muscle.
-- Lactate is released from the muscles to the
blood and travels to the liver for conversion to
pyruvate and, ultimately to glucose.
CORI’S CYCLE
SUBSTRATES FOR GLUCONEOGENESIS
Glycogenic AA
SUBSTRATES FOR GLUCONEOGENESIS
Glycerol
SUBSTRATES FOR GLUCONEOGENESIS
Propionate – In ruminants
Bypass number 1.
Pyruvate to phosphoenolpyruvate.
This bypasses pyruvate kinase.
Step 1 : pyruvate to oxaloacetate
Enzyme = pyruvate carboxylase
located inside mitochondria.
Sub reactions are :
Enz-biotin + ATP + HCO3- Enz-carboxybiotin +
ADP + Pi
Enz-Carboxybiotin + pyruvate Enz-biotin +
oxaloacetate
The overall reaction is
pyruvate + HCO3- + ATP oxaloacetate + ADP + Pi
For gluconeogenesis, oxaloacetate must leave
the mitochondria because all the rest of the
gluconeogenesis enzymes are in the cytosol.
mitochondrial membranes are nearly
impermeable to oxaloacetate.
So how does it get out?
Transport - three steps
Conversion of OAA to Malate(Mitochondria)
NADH+H+ NAD+
Oxaloacetate Malate
Malate Dehydrogenase
malate can be transported through the
mitochondrial membrane into the cytosol
In cytosol, oxaloacetate is regenerated from
malate by malate dehydrogenase – cytosolic
enzyme
NAD+ NADH+H+
Malate Oxaloacetate
Malate Dehydrogenase
Step 1 : Decarboxylation of OAA to PEP
Enzyme = PEP carboxykinase cytosolic enzyme
oxaloacetate + GTP phosphoenolpyruvate
+ CO2 + GDP
uses GTP, not ATP.
CO2 added is lost in this step.
NET so far:
pyruvate + ATP + GTP PEP + ADP + GDP + Pi
High cost = two energy rich phosphates
so a total of four high energy bonds are
already utilized here per glucose to be
synthesized.
Then the remaining steps are reversible
reactions using glycolytic enzymes in steps
upto formation of fructose-1,6-bisphosphate
Bypass number 2. Fructose-1,6-
bisphosphate to fructose-6-phosphate
Enzyme = fructose-1,6-bisphosphatase
Reaction:
fru-1,6-bisphosphate + H2O fructose-6-P + Pi
bypasses phosphofructokinase
a simple hydrolysis, releasing Pi
highly exergonic, irreversible
enzyme is highly regulated
F6P isomerizes to glucose-6-phosphate via
phosphoglucoisomerase
Bypass number 3.
Glucose-6-phosphate to glucose.
Enzyme = glucose-6-phosphatase (luminal
surface of ER membrane)
Reaction:
glucose-6-phosphate + H2O glucose + Pi
bypasses hexokinase
highly exergonic, irreversible
not present in muscle
REGULATION:
a) Availability of substrates
b) Activity or amount of key enzymes of
glycolysis and gluconeogenesis.
Substrate availability:
a) Lactate is produced in muscle during exercise
b) Amino acids are released from muscle during
fasting and intake of high protein diet
increases gluconeogenesis
c) Glycerol is released from adipose tissue
(lipolysis) when insulin is low and glucagon is
high
REGULATION OF KEY ENZYMES:
1. PYRUVATE CARBOXYLASE.
activated by acetyl-CoA (required) vs. pyruvate
kinase, inhibited by acetyl CoA.
high levels of acetyl-CoA signals that enough
carbon substrate available for citric acid cycle.
It is inhibited by ADP
2. FRUCTOSE 1,6 BISPHOSPHATASE
Strongly inhibited by AMP, low energy
Strongly inhibited by fructose-2,6-bisphosphate,
high glucose
Recall that the reciprocal enzyme,
phosphofructokinase, in glycolysis, is strongly
activated by AMP and fructose-2,6-bisphosphate.
Fructose-2,6-bisphosphate is the most important
regulator of glycolysis and gluconeogenesis
through its reciprocal effects on
fructose 1,6-bisphosphatase and
phosphofructokinase.
Glucagon: hormone released when glucose levels
are low.
signal to elevate blood glucose levels
increases intracellular levels of cAMP in liver
and elsewhere
cAMP activates protein kinase A which in
turn activates cAMP-dependent protein kinases
stimulating gluconeogenesis and
glycogenolysis
Insulin
-- High levels of glucose induce release of insulin
from β-cells of islets of Langerhan in the pancreas.
-- Insulin is polypeptide hormone.
-- Detected by receptors at surface of muscle cells.
-- Increases glycogenesis in muscle.
-- Intracellular signal pathway involves complex
sequential phosphorylations and
dephosphorylations.
Summary of Gluconeogenesis
purpose- alternative source of glucose rather
than dietary carbohydrates or glycogen
breakdown
primary precursors are pyruvate, lactate,
glycerol, part of fatty acids and certain amino
acids (glucogenic)
3 essentially irreversible steps of glycolysis
are bypassed
regulated via pyruvate carboxylase,
fructose 1,6 bisphosphatase, and
phosphofructokinase-2/fructose
bisphosphatase-2
