Gluconeogenesis

1. Gluconeogenesis Dr. Sooad Al-Daihan Biochemistry department

2. Introduction • Some tissues, such as the brain, and red blood cells require a continuous supply of glucose as a metabolic fuel. • During a prolonged fast, hepatic glycogen stores are depleted, and glucose is formed from precursors such as lactate, pyruvate, glycerol, andα-ketoacidsby a special pathway, gluconeogenesis, that requires both mitochondrial and cytosolic enzymes.

4. Gluconeogenesisoccurs mainly in liver. • It also occurs to a more limited extent in kidney & small intestine under some conditions.  • Synthesis of glucose from pyruvate utilizes many of the same enzymes as Glycolysis. • Almost reverse of glycolysis exceptfor 3 reactions ,which are essentially irreversible. • Hexokinase (or Glucokinase) • Phosphofructokinase • Pyruvate Kinase. • These steps must be bypassed in Gluconeogenesis.

5. Four enzymes are needed to reverse the 3 irreversible steps of glycolysis: • Mitochondrial ‐ Pyruvate Carboxylase (liver, kidney but not in muscle) • Cytoplasmic ‐ Phosphoenolpyruvate (PEP) Carboxykinase • Cytoplasmic ‐Fructose‐1,6,‐Bisphosphatase • Cytoplasmic ‐Glucose 6‐Phosphatase

6. 1-Bypass of Pyruvate Kinase: In gluconeogenesis PK is bypassed by 2 enzyme catalyzed reactions: 1- Pyruvate Carboxylase : Pyruvate is carboxylated to oxaloacetate in the mitochondria . pyruvate + HCO3- + ATP  oxaloacetate + ADP + Pi 2- PEP Carboxykinase : Oxaloacetate is decarboxylated and phosphorylated to yield PEP in the cytosol . oxaloacetate + GTP  PEP + GDP + CO2

7. Transport of oxaloacetate to the cytosol

8. 2- Bypass of Phosphofructokinase: In gluconeogenesis PFK bypassed by Fructose 1,6 ‐ bisphosphatase reaction (Removes phosphate group) fructose‐1,6‐bisP + H2O fructose‐6‐P + Pi 3- Bypass of Hexokinase (or Glucokinase) In gluconeogenesis this reaction bypassed by glucose 6‐phosphatase reaction (Removes phosphate group) : Glucose-6‐P + H2O Glucose+ Pi Free glucose is formed by the action of glucose‐6‐ phosphatase in liver and kidney while it is absent in muscles and adipose tissues Glucose can not be formed by these organs

9. Total Energy Cost

10. Precursors for Gluconeogenesis i- Cori Cycle • Lactate released by active skeletal muscle or red blood cells is carried to the liver where it is converted to glucose by gluconeogenic pathway (Coricycle) and released for reuptake by skeletal muscle ii- Glucose-Alanine Cycle • Protein broken down in skeletal muscle during exercise • Amino acids converted to alanine and released by skeletal muscle • Taken up by liver and converted to glucose and released for reuptake by skeletal muscle

11. iii- Glycerol • Glycerol released from adipocytes and skeletal muscle during lipolysis. •  Glycerol enters gluconeogenic pathway as dihydroxyacetone phosphate “active form of glycerol”. •  2 glycerol required to make one glucose in liver and kidney in fasting or low CHO diet . • Glycerol cannot be utilized in adipose tissue due to the lack of glycerol kinase in the adipose tissue.

12. iv- Propionate: • Propionyl‐CoA is converted to the TCA intermediate, succinylCoA. • This conversion is carried out by the ATP‐requiring enzyme, propionyl‐CoA carboxylase then methylmalonyl‐CoA epimerase and finally the vitamin B12 requiring enzyme, methylmalonyl‐CoA mutase • The utilization of propionate in gluconeogenesis only has quantitative significance in ruminants.

13. Regulation of Gluconeogenesis • Control of glycolysis and gluconeogenesis is reciprocal • Gluconeogenesis and Glycolysis are regulated by similar effector molecules but in the opposite direction • When one pathway is activated , the other is inhibited • Gluconeogenesis is subject to both: - Hormonal control by Glucagon, Cortisol, Adrenaline and Insulin - Allostericregulation of gluconeogenic enzymes

14. Hormonal Regulation Glucagon, Cortisol, Adrenaline • Are secreted during fasting, stress and sever muscular exercise • Inducegluconeogenic enzymes • Repress glycolytic enzymes Insulin • Secreted after CHO meal • Induceglycolytic enzymes • Repress gluconeogenicenzymes

15. Allosteric regulation

16. Uronic Acid Pathway Dr. Sooad Al-Daihan Biochemistry department

17. Overview • It is an alternative oxidative pathway for glucose that doesn’t lead to ATP generation. • It includes oxidation of glucose to • Glucuronic acid • Ascorbic acid • It occurs mainly in the liver cytoplasm.

18. Metabolic reactions • Glucose 6-phosphate is isomerized to glucose 1-phosphate • Glucose 1-phosphate reacts with uridine triphosphate (UTP) to form uridinediphosphate glucose (UDPGlc) in a reaction catalyzed by UDPGlcpyrophosphorylase • UDPGlcis oxidized at carbon 6 by NAD-dependent UDPGlc dehydrogenase in a two-step reaction to yield UDP-glucuronate 1 2 3

19. UDP Glucuronic acid (active form) is needed in: • Conjugation to less polar compounds as bilirubin, steroids and some drugs making them more water soluble (detoxicated) . • Synthesis of glycosaminoglycans (mucopolysaccharide) as heparin, hyaluronic acid. • In plants and some animals (not Human) glucuronic acid serves as a precursor of L-ascorbic acid. • The uronic acid pathway also provides a mechanism by which dietary D-xylulose enter the central pathway.