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GLUCONEOGENESIS
Dr. Catherine L. Co-Reportoso | October 31, 2018 LE3 TRANS2
L.E. # 3 - Trans # 2 Group F: Ramirez, R.J., Rapisora, Raymundo, Regalado, Reyes, A.A. 1 of 10
Breakdown of 1 mol glucose o 1 FADH2 x 1.5
o Aerobic - 2 mol pyruvate, yields 5-7 ATP depending on o 1 ATP (GTP)
the shuttle system used Site for entry of most amino acids for gluconeogenesis
o Anaerobic - 2 mol lactate, yields 2 ATP
Glycolysis is linear pathway consisting of 10 total steps Recall:
ATP is consumed in the 1st and 3rd reactions within the first Anaplerotic reactions – replenish TCA intermediates
phase (energy investment phase) o Pyruvate carboxylase (pyruvate to oxaloacetate)
ATP is produced in the 7th (Phosphoglycerate kinase) and Cataplerotic reactions - utilizes TCA intermediates
10th reactions (Pyruvate kinase) within the second phase o Helps remove buildup of excess molecules
(energy generation phase). o Intermediates can be used for biosynthesis of other
Function in fuel metabolism via provision of ATP macromolecules
Steps 1,3, and 10 are irreversible o Malate to aspartate during Malate-Aspartate shuttle
o Step 1 (Hexokinase / Glucokinase): dedicates glucose [2021 A Trans]
to the cell for metabolism
o Step 3 (PFK-1): rate-limiting step II. GLUCONEOGENESIS
Step 7 is a reversible step GLUCONEOGENESIS
Activated during fed state [2021 A Trans] A. DEFINITION
First step
o Glucose Glucose 6-Phosphate The process of synthesizing glucose or glycogen from
o Enzyme: Hexokinase/Glucokinase (Glucokinase has noncarbohydrate precursors [Harper]
higher Km than hexokinase for glucose) Synthesis of glucose from non-carbohydrate compounds
o Phosphate: comes from ATP (Investment phase) Converts 2 moles of pyruvate to 1 mole of glucose
Substrate level of phosphorylation: Phosphoglycerate Also involves TCA cycle intermediates especially
kinase and pyruvate kinase because they have two high oxaloacetate
phosphate energy compounds. [Lecturer] Gluconeogenic flux is regulated primarily by fructose-2,6
1, 3 Biphosphoglycerate and Phosphoenol pyruvate donate biphosphate
phosphate to ADP to become ATP. They have higher - ∆G. Energetically expensive (Consumes 6 ATP when
[Lecturer]
converting 2 mol of pyruvate to 1 glucose)
The reverse of Gluconeogenesis. Because they share the
same enzymes on the reversible reactions. In the irreversible Functions
reactions, these are the steps bypassed in gluconeogenesis: Enables maintenance of blood glucose levels long after all
o Reactions catalyzed by Pyruvate kinase, PFK1 and dietary glucose has been absorbed and completely
Glucokinase/Hexokinase oxidized and glucose stored as glycogen has been used up
[Devlin]
C. TRICARBOXYLIC ACID CYCLE Maintain level of intermediates of citric acid cycle [2021 A]
T Recycles waste products of metabolite [2021A]:
→ Lactate from: Muscle and Erythrocytes
→ Glycerol from: adipose tissues
Locations
Major gluconeogenic tissues: liver and kidney
o Overnight fast
Liver: 90%
Kidney: 10%
o Prolonged fasting
Kidney: 40%
Already occurs in lesser extent in the liver 4 hours after meal
o Not major source compared to glycogenolysis
Renal gluconeogenesis occurs about 2 days of fasting
Gluconeogenesis is highly active during 2-5 days of fasting
After 5 days, the body the amino acids are conserved
while the body undergoes ketogenesis [2021A]
Occurs largely in the CYTOSOL
Major substrates
Glucogenic Amino Acids
Lactate
Glycerol
Propionate [Harper]
B. STEPS, REACTIONS AND PRODUCTS
7 of the 10 enzymatic reactions of gluconeogenesis are the
reverse of glycolytic reactions [Lehninger].
→These reactions have a 𝚫G near 0
Figure 5. Diagram of Kreb’s Cycle [Quora]
→The 3 irreversible steps are bypassed by a separate set of
Final common pathway for oxidation of CHO, lipids, and enzymes that are also exergonic:
proteins with Acetyl CoA as substrate. Carboxylation of pyruvate to PEP
Occurs in the mitochondria Conversion of Fructose-1,6-bisphosphate to fructose-
Occurs at inner mitochondria and accounts for the two-thirds 6-phosphate
of ATP produced in fuel oxidation. Glucose-6-phosphate to Glucose
10 ATP per cycle
o 3 NADH x 2.5
Biochemistry Gluconeogenesis 2 of 10
11. F6P will be isomerized to glucose-6-phosphate (G6P) using
phosphohexose isomerase
12. G6P will be dephosphorylated by glucose-6-phosphatase to
yield one glucose molecule
Steps
*Remember that 2 mol of pyruvate are needed to generate 1 mol Alternative route
of glucose • OAA is reduced to malate in the mitochondria
catalyzed by malate dehydrogenase
A. PYRUVATE TO PHOSPHOENOL PYRUVATE (PEP)
• Malate is transferred to the cytosol
• Malate is then oxidized back to OAA in the cytosol
catalyzed by malate dehydrogenase
• Cytosolic PEPCK will catalyze the conversion of OAA to
PEP
Biochemistry Gluconeogenesis 4 of 10
Common enzyme during anaerobic glycolysis o In the liver, alanine undergoes transamination via alanine
Catalyzes the reversible reaction of converting lactate to pyruvic aminotransferase to produce pyruvate
acid o Amino group is donated to -ketoglutarate and becomes
Redox reaction glumatate
NAD+ is converted to NADH • Cys, Gly, Thr, Ser and Trp also enters pyruvate [Lecturer’s PPT]
The cycling of lactate and glucose between liver and peripheral • Pyruvate enters gluconeogenesis
tissues is called the Cori Cycle (Glucose Lactate Cycle)
Entry through TCA
B. GLUCOGENIC AMINO ACIDS All glucogenic AAs that participates in TCA cycle will enter
Amino Acids that can be converted into glucose through gluconeogenesis via oxaloacetate
gluconeogenesis [2021 B Trans] Consumes 4 ATP
Carbon atoms of AAs derived from proteins can be catabolized o Pyruvate carboxylase reaction is skipped
to pyruvate or to intermediates of the citric acid cycle [Lehninger] o Since 2 moles of Oxaloacetate are also used for
Alanine and Glutamine are important glucogenic AAs in gluconeogenesis 2 ATP is deducted
mammals that transport amino groups from extrahepatic tissues Anaplerotic reactions are the reactions that lead to net
to the liver. synthesis of TCA cycle intermediates and support
Amino group is first removed in the liver mitochondria and the gluconeogenesis as they provide for net synthesis of
remaining carbon skeletons are funneled into gluconeogenesis Oxaloacetate.
[Lehninger]
o Examples are reactions catalyzed by pyruvate carboxylase
All amino acids except Leucine and Lysine can supply carbon and glutamate dehydrogenase
for net synthesis of glucose by gluconeogenesis [Devlin] Reactions catalyzed by glumatate-oxaloacetate
aminotransferase are not anaplerotic as net synthesis of a TCA
cycle intermediate is not accomplished.
Glucogenic AAs enter through 4 intermediates:
-ketoglutarate
→Succinyl CoA
→Fumarate
→Oxaloacetate
C. GLYCEROL
Can be derived from the degradation of Triacylglycerides
(TAG) by lipases.
Hydrolysis of TAGs yield three fatty acids and glycerol [Devlin]
Results to the formation of the glycerol molecule along with
three molecules of fatty acid chains.
Glycerol is converted to an intermediate of glycolysis,
Dihdroxyacetone phosphate [Lecturer]
Glycerol enters the gluconeogenesis while the remaining fatty
acids undergo beta oxidation to produce acetyl-CoA
Consumes: 2 ATP if all carbon of glucose is derived from
glycerol
’
Figure 13. Glucogenic Amino Acids According to Site of Entry [Lehninger]
*Are also ketogenic amino acids
Phosphorylation of Glycerol
Carbon 3 of glycerol is phosphorylated
Enzyme: Glycerol kinase
Produces Glycerol-3-phosphate
Requires ATP as source of phosphate group
Adipocytes lack glycerol kinase thus they cannot directly
phosphorylate glycerol [Lehninger]
→ Glycerol produced in adipocytes is transported to the liver.
Biochemistry Gluconeogenesis 6 of 10
The Cori Cycle uses Lactate as a Substrate and undergoes 2 moles of ATP are produced along with 2 pyruvate and 2
anaerobic glycolysis to produce 2 ATP NADH
→Occurs during exercise when there is high production of NADH produced will not go to ETC and instead will be
lactate when O2 supply is depleted thus cannot proceed to ETC. oxidized by LDH to convert Pyruvate to Lactate
→Reoxidizes NADH to NAD+ Lactate is then transported to the major gluconeogenic
→Less ATP production in muscle (only 2 ATP) organs [2021 A Trans]
In the liver, lactate undergoes gluconeogenesis via pyruvate
there is a consumption of 6 ATP. During Gluconeogenic Step
The Cahill Cycle utilizes Alanine produced in the muscles Hepatic LDH oxidizes lactate to reform pyruvate which transfers
through the transamination of pyruvate. free electron to NAD forming NADH
→ In contrast to the Cori cyle, it undergoes aerobic
Pyruvate then proceeds to steps of gluconeogenesis
glycolysis which is more energy efficient for muscles
[Devlin]
Presents liver with amino nitrogen (ammonium) which must be
disposed as urea [Devlin]
→ In the liver, the urea cycle consumes 4 ATP while
gluconeogenesis consumes 6 ATP, total consumption of 10
ATP
During Transamination
Close relationship between urea synthesis and glucose
synthesis as gluconeogenesis from AAs imposes nitrogen
load on liver [Devlin]
Muscles degrade amino acids for energy needs and the
resulting nitrogen is transaminated to pyruvate to form alanine
Alanine undergoes transamination again to form pyruvate
→ Alanine transaminase (ALT) converts glutamate into -
ketoglutarate and vice versa
→ ALT is formerly known as serum-glutamate pyruvate
transaminase (SGPT) [Lecturer]
Resulting alanine is shuttled to the liver where nitrogen enters
the urea cycle (4 ATP) and pyruvate is utilized to make
glucose (6 ATP)
The cycle utilizes 10 ATPs and produces 5/7 ATPs Figure 23. Reciprocal Regulation of Gluconeogenesis [Lecturer’s PPT]
ATP Produced: C. COVALENT MODIFICATION
→ 2 ATP + 2 NADH Due to reversible phosphorylation
→2 NADH x 1.5 =3 ATP (Glycerol-Phosphate Shuttle) Rapid
→ 2 NADH x 2.5 =5 ATP (Malate-Aspartate Shuttle) Glucagon and Epinephrine
o Inhibits glycolysis and stimulates gluconeogenesis
o Increases concentration of cAMP
o Activates cAMP-dependent protein kinase that leads to
inactivation of pyruvate kinase
o Affects the concentration of fructose 2,6-bisphosphate
Glucagon other functions[Lippincott]:
o Changes in allosteric effectors - lowers the level of
fructose 2,6-bisphosphate, resulting in activation of fructose
1,6-bis-phosphatase and inhibition of phosphofructokinase-
1, thus, favoring gluconeogenesis over glycolysis
o Induction of enzyme synthesis - increases the
transcription of the gene for PEP-carboxykinase, thereby
increasing the availability of this enzyme as levels of its
substrate rise during fasting
Figure 22. Conversion of Alanine to -ketoglutarate and vice versa o Covalent modification - Glucagon binds its G protein-
[Lecturer’s PPT] coupled receptor and, via an elevation in cyclic AMP
(cAMP) level and cAMP-dependent protein kinase activity,
stimulates the conversion of hepatic pyruvate kinase to its
inactive (phosphorylated) form. This decreases the
conversion of PEP to pyruvate, which has the effect of
diverting PEP to the synthesis of glucose.
VI.REGULATION OF GLUCONEOGENESIS
A. RECIPROCAL REGULATION
Glycolysis and gluconeogenesis share the same pathway but in
opposite directions. [Harper’s]
Both pathways are reciprocally regulated
Biochemistry Gluconeogenesis 8 of 10
Figure 24. Covalent modification of Glucagon [2021 A Trans] Phosphofructokinase-1
B. RATE OF ENZYME SYNTHESIS Inhibited by citrate
Induction and repression of key enzymes take hours. Activated by AMP
Enzymes involved catalyze physiologically irreversible non- 90% inhibited at normal levels of ATP
equilibrium reactions. when ATP is converted to ADP for energy, AMP increases
Enzymes of gluconeogenesis have low activity when there is AMP acts as an indicator for the energy status of the cell
super-fluidity of glucose. AMP increases glycogenolysis activity by activation glycogen
Enzymes that induce gluconeogenesis, glucocorticoids and phosphorylase
glucagon-stimulated cAMP, are antagonized by the presence of Inhibition of PFK-1 by ATP = increase in glucose-6-phsophate
insulin. Increased G6P = inhibits glucose uptake and action of
hexokinase
Table 5. Regulatory and Adaptive Enzymes Associated with Carbohydrate
Metabolism (Table 19-1 in Harper’s) Fructose 2,6-Bisphosphate
Inducer Repressor Activator Inhibitor Potent positive allosteric activator of PFK-1, by increasing
Glycogenolysis, glycolysis, and pyruvate carboxylation – The PFK-1 affinity for fructose-6-phosphate
following enzymes increases in Carbohydrate feeding and Inhibitor of fructose 1,6-bisphosphate by increasing fructose
decreases in Fasting & Diabetes 1,6-bisphosphate’s Km
Glycogen Insulin, Glucagon
Synthase glucose-6-
phosphate
Hexokinase Glucose-
6-
Phosphate
D. ALLOSTERIC EFFECTORS
Acetyl-CoA
An allosteric activator of pyruvate carboxylase
Together with pyruvate carboxylase, catalyzes the synthesis of
oxaloacetate from pyruvate
As acetyl-CoA is formed from pyruvate, this ensures production
of oxaloacetate
Activation of pyruvate carboxylase = inhibition of pyruvate
dehydrogenase
Biochemistry Gluconeogenesis 9 of 10
VII. PHYSIOLOGICAL IMPORTANCE OF 4. An increased NAD/NADH ratio in alcohol metabolism
GLUCONEOGENESIS inhibits gluconeogenesis by
Some of the organs in the body requires continues supply of a. Favoring malate Oxaloacteate
glucose b. Favoring pyruvate lactate
o These organs are supplied by a non-carbohydrate source c. Inhibit DHAP glycerol 3P
in the process of gluconeogenesis. [Lecturer] d. Inhibit alanine pyruvate
o These organs are the brain, heart, RBC, renal medulla,
cornea and testis [2021 A Trans] 5. Glycerol from TAG degradation enters gluconeogenesis
Four gluconeogenic sources: via which intermediate?
o Lactate a. DHAP
o Glucogenic amino acids b. Glyceraldehyde 3P
o Glycerol c. Fructose 1,6 bisP
o Propionyl CoA d. Pyruvate
Two Major Gluconeogenic Organs:
o Liver 6. Which reaction in gluconeogenesis utilizes energy?
o Kidneys a. Oxaloacetate PEP
b. Glucose 6P Glucose
• Only two because of Glucose 6 - phosphatase (the
c. Fructose 1,6bisP Fructose 6P
enzyme that converts glucose 6 phosphate to glucose).
d. 1,3Phosphoglycerate Glyceraldehyde 3P
Glucose 6 - phosphatase is only found in the endoplasmic
reticulum of liver and kidneys [Lecturer]
7. In the glucose synthesis via Cori cycle and Glu-Ala cycle
Source of glucose during fasting [2021B Trans] respectively, what is the correct number of ATP pair used?
o 10-18 hours of fasting: glycogenolysis a. 4 & 6
o Prolonged fasting: Gluconeogenesis b. 6 & 4
CHO Metabolism pathways during fed state [2021B Trans]: c. 6 & 10
o Glycolysis d. 10 & 6
o Glycogenesis
Effect of Alcohol Intake Answers: 1. c., 2. a. 3. b., 4. b. 5. a., 6. a., 7. c.
IX. REFERENCES
Hypoglycemia
o caused by the consumption of alcohol and drinking Devlin, T. (2011). Textbook of Biochemistry with Clinical Correlations (7th ed).
United States, John Wiley & Sons, Inc.
alcohol after strenuous exercise. Rodwell, V. et al, (2003). Harper’s Illustrated BIochemistry (30th ed). McGraw Hill
o results from the inhibitory effects of alcohol on hepatic Education.
gluconeogenesis and occurs under circumstances of Nelson, D. and Cox, M. (2008) Lehininger Principles of Biochemistry (5th ed) New
York, W.H. Freeman Company
hepatic glycogen depletion [Devlin]. Harvey R., and Ferrier D., (2014) Lippincot’s Illustrated Reviews (6th ed).
During Alcoholic intoxication: Philadelphia, Wolters Kluwer/Lippincott Williams and Wilkins
o There’s an increase in NADH/NAD ratio that inhibits Dr. Reportoso’s Lecture PPT
2021 A and 2021 B Trans
gluconeogenesis
o NADH is used by lactate dehydrogenase to convert
pyruvate to lactate
o Triacylglycerols are increased because of NADH/NAD
ratio.
The DHAP will be converted to glycerol 3 –
phosphate and later to glycerol.
Glycerol is the backbone of TAGs.
VIII.REVIEW QUESTIONS
1. Glucogenic amino acids enter which Krebs cycle
intermediate for glucose synthesis?
a. Citrate
b. Malate
c. Oxaloacetate
d. Succinate
Biochemistry Gluconeogenesis 10 of 10