Glycolysis Introduction to Metabolism Regulation of Metabolism Overview of Glycolysis Reactions of Glycolysis

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1 Glycolysis Introduction to Metabolism Regulation of Metabolism Overview of Glycolysis Reactions of Glycolysis Suggested Reading: Lippincot s Ilustrated reviews: Biochemistry

2 Glycolysis, an example of metabolic pathway The product of one reaction is the substrate of the next reaction

3 Metabolic pathways intersect to form network of chemical reactions

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6 Regulation of Metabolism Signals from within the cell Substrate availability, product inhibition, allosteric Rapid response, moment to moment Communication between cells (intercellular) Slower response, longer range integration Second messenger Ca 2+ / phosphatidylinositol system Adenyl cyclase system

7 Communication between cells; Commonly used mechanisms

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10 INTRACELLULAR EFEECTS Activated enzymes Inhibited Enzymes cell s ion channels bind to promoter

11 GLYCOLYSIS Universal Pathway: In all cell types Generation of ATP With or without O 2 Anabolics Pathway: biosynthetic precursors

12 The Two Phases of the glycolytic Pathway Preparative Phase ATP-generating Phase 1 Glucose (C6) 1 ATP 1 ATP 1 Fructose 1,6 bisphosphate (C6) 2 Triose Phosphate (C3) 2 NADH 2 ATP 2 ATP 2 Pyruvate (C3)

13 Glycolysis occurs in all human cells Glucose Pyruvate acetyl CoA No O 2 requirement for glycolytic anaerobic fermentation Lactate CO2 TCA CO2 O 2 requirement for PDH & TCA activities

14 Brain Tissues with an Absolute or high Requirement for Glucose Red Blood Cells Cornea lens and retina Kidney Medulla, Testis Leukocytes White muscle fibers

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16 Hexokinase Glucokinase Occurance In all tissues In liver Km < 0.02 mm mm Specificity Glc., Fruc, Man, Gal Glc. induction Not induced insulin, Glc Function At any glucose level Only > 100 mg/dl

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22 Mechanism of the reaction: E and S form covalent linkage S is oxidized and NADH is formed NADH is released P i attacks the thioester bond releasing the product

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25 -ATP -ATP 2 NADH 2 ATP Is Oxygen needed? 2ATP

26 Synthesis of 2,3 bisphosphoglycerate in RBC Oxygen delivery to tissues

27 Pyruvate + NADH Lactate + NAD+

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31 Lactate Production Cells with low energy demand To cope with increased energy demand in rigorously exercising muscle Hypoxia lactate level is increased 5 to 10 folds to survive brief episodes of hypoxia

32 ph of the plasma Lactic Acidosis The most common cause of metabolic acidosis Production of lactic acid utilization of lactic acid Pyruvate + NADH Lactate + NAD + Most common cause: Impairment of oxidative metabolism due to collapse of circulatory system. Impaired O 2 transport Respiratory failure Uncontrolled hemorrhage

33 Lactic Acidosis Direct inhibition of oxidative phosphorylation Hypoxia in any tissue Alcohol intoxication ( high NADH/ NAD+ ) Gluconeogenesis Pyruvate Dehydorgenase TCA cycle activity Pyruvate carboxylase

34 Inorganic Inhibitors of Glycolysis Fluoride Fluoride inhibits Enolase Fluoridated water Prevention of Dental Carries bacterial enolase

35 Inorganic Inhibitors of Glycolysis Arsenic Poisoning Pentavalent Arsenic (Arsenate) competes with phosphate as as a substrate for GA3PDH ATP synthesis Trivalent Arsenic (Arsenite) Forms stable complex with -SH of lipoic acid Pyruvate Dehydrogenase α ketoglutarate Dehydrogenase Neurological dissturbances.death

36 Pyruvate Kinase Deficiency The most common among glycolytic enzyme deficiencies ( 95% of cases PK ; 4% PGI ) RBC s are affected Mild to severe chronic hemolytic anemia ATP is needed for Na+/K+ pump maintain the flexible shape of the cell Low ATP premature death of RBC Abnormal enzyme; mostly altered kinetic properties

37 Alterations observed with various mutant forms of pyruvate kinase

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42 Regulation of PFK by Fructose 2,6- bisphosphate Fruc. 6-phosphate + ATP PFK-2 Fruc. 2,6 bisphosphate + ADP

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44 Regulation by ATP and AMP; why AMP ADP + ADP ATP + AMP

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49 Next topic: Gluconeogenesis

50 Energy-yielding nutrients Complex molecules C A T A B O L I M Energy-poor end products A N A B O L I S M Precursor molecules

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