Lecture 20 Gluconeogenesis

Basics
Example of anabolic pathway
Comparison with a parallel catabolic pathway (glycolysis)
appears to be reverse of glycolysis
Commonality
7 reactions of glycolysis

Difference
3 reactions of glycolysis that are highly exergonic
4 reactions of gluconeogenesis
Pyruvate -> Oxaloacetate
Oxaloacetate -> PEP
Fructose 1,6-P2 -> Fructose 6-P
Glucose 6-P -> Glucose

Reason for difference
Gluconeogenesis is endergonic
Different enzymes are required for separate regulation of the two pathways

Physiological role
importance of glucose availability for human body
brain uses most of glucose daily (160 g / day)

amino acids. glycerol.Body fluid carries 20 g. and TCA cycle intermediates 4 Unique Reactions Pyruvate -> Oxaloacetate Substrates and Products Pyruvate + Bicarbonate + ATP -> Oxaloacetate + ADP + Pi Direction: Driven forward with ATP energy Enzyme: Pyruvate Carboxylase Mechanism Biotin as carboxyl group carrier Biotin linked to a lysine residue ATP activates bicarbonate Bicarbonate transferred to Biotin Regulation Allosteric Activator Acetyl-CoA and ATP when acetyl-CoA or ATP is low . or DHAP lactate. Oxaloacetate. ~200 g available as glycogen Backup supply of glucose compare with fat: catabolism does not generate glucose Possible precursors of gluconeogenesis: enter at Pyruvate.

6-P2 -> Fructose 6-P Substrates and Products Fructose 1.6-P2 + H2O -> Fructose 6-P + Pi Direction: driven forward .glycolysis->TCA cycle is activated gluconeogenesis is inhibited Location Mitochondrion Compartmentalization helps preventing futile cycle of simultaneous glycolysis and gluconeogenesis Transport of Oxaloacetate from Mitochondria to Cytoplasm Conversion to malate and back to Oxaloacetate Oxaloacetate -> Malate: reverse of TCA cycle reaction through malate dehydrogenase Coupled with NADH -> NAD+ and NAD+ -> NADH Oxaloacetate -> PEP Substrates and Products Oxaloacetate + GTP -> PEP + GDP + CO2 Direction: driven forward to PEP production Energetically favorable GTP energy used Equivalent of using ATP because ATP is used to regenerate GTP from GDP Enzyme: PEP carboxykinase (PEPCK) Fructose 1.

6-BPase activated use ATP cAMP-dependent protein kinase activated by glucagon Example of Substrate Cycle Only one of the two anti-parallel reactions is turned on at a time Compare Fasting State Fed State .6-BPase Two enzyme activities are in the same protein! "Bifunctional" or "Tandem" enzyme Allosteric regulation: F-6-P PFK-2 activated F-2.6-bisphosphate generated by PFK-2 enzyme Different from PFK (or PFK-1) reversed to F-6-P by Fructose-2.6-bisphosphatase (F-2.6-BPase) PFK-2 vs F-2.6-BPase inhibited Regulation by phosphorylation: cAMP-dependent protein kinase PFK-2 inhibited F-2.regulation activation citrate inhibition fructose 2.hydrolysis of phosphate ester is thermodynamically favorable Enzyme: Fructose 1.6-bisphosphatase .

T3 T1: G-6-P from cytosol to ER T2: Glucose from ER to cytosol T3: Pi from ER to cytosol .AMP synergistic Opposite to PFK regulation Glucose 6-P -> Glucose Substrates and Products Glucose 6-P -> Glucose + Pi Direction: driven forward exergonic due to phosphate ester hydrolysis Enzyme: glucose 6-phosphatase Body Location: absent from muscle and brain . T2.No glucose produced through gluconeogenesis present in liver and kidney Cellular location: Endoplasmic Reticulum Transporters: T1.

GLUT2: Glucose from cytosol to extracelluar space mechanism Histidine residue phosphorylated by G-6-P Water releases Pi from histidine regulation substrate-level control Km is higher than normal range of substrate concentration (G-6-P) Low Carb diet or Starvation (ref: Chapter 23) Body switches from glucose to fat as energy source generation of ketone bodies Ketosis Effect on weight loss initially water is a big part of weight loss glycogen is associated with 3X weight of water Health risk of Low Carb diet increased intake of fat -> heart disease reduced intake of fruit -> cancer Whole body Physiology in humans Cori cycle Lactate -> pyruvate -> glucose Redistribution of lactate and glucose .

22) reciprocal control between glycolysis and gluconeogenesis Depends on glucose availability as well as energy status Energy status: AMP/ATP low energy glycolysis UP provide ATP .Lactate buildup in muscle Shortage of oxygen NAD+ cannot be regenerated through cellular respiration NADH -> NAD+ by reducing pyruvate to lactate Lactate transported to liver Lactate reoxidized to pyruvate pyruvate can be converted to glucose Liver shares metabolic burden of exercise Summary and comparison with Glycolysis Overall reaction Six ATP equivalents used 4 ATP 2 GTP 2 pyruvate converted to glucose Compare with energy chart of 11 reactions (Fig 18.

high energy gluconeogenesis UP use energy to produce glucose and glycogen Substrate-level control Glucose-6-Phosphatase Allosteric control Acetyl-CoA Citrate F-2.6-P2 ATP/AMP .