The matrix contains Pyruvate Dehydrogenase, enzymes of Krebs Cycle, and other pathways, e.g., fatty acid oxidation & amino acid metabolism.

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1 Glycolysis occurs in the cytosol of cells. matrix Pyruvate enters the mitochondrion to be metabolized further. cristae intermembrane space Mitochondrial Compartments: inner membrane mitochondrion outer membrane The matrix contains Pyruvate Dehydrogenase, enzymes of Krebs Cycle, and other pathways, e.g., fatty acid oxidation & amino acid metabolism. 1

2 2 Citric acid cycle Krebs cycle, tricarboxylic acid cycle TCA The central function is the oxidation of acetyl CoA to CO2 - It is the final common pathway for oxidation of fuel molecules - Acetyl CoA is derived from the metabolism of fuel molecules as amino acids, fatty acids, and carbohydrates. - Citric acid cycle is also an important source of precursors Some intermediates are precursors of amino acid One of the intermediates is used in the synthesis of porphorins Another is used in the synthesis of fatty acids and sterols. - Citric Acid Cycle located in the mitochondrial matrix

3 3

4 Citric acid cycle is also an important source of precursors for biosynthetic reactions 4

5 Citric acid cycle intermediates are always in flux 5

6 The Citric acid cycle - Citric acid cycle contains a series of oxidation-reduction reactions - Carbon entering the cycle, leaves fully oxidized as CO2. - High energy electrons leave the cycle with high energy electron carriers as NADH and FADH2. - Very little ATP is made directly in the cycle. - No oxygen is used in the cycle. Overall reaction 3NAD+ + FAD + GDP + Pi + 2H2O + acetyl-coa 3NADH + FADH2 + GTP + CoA + 2CO 2 + 3H+ 6

7 The citric acid cycle oxidizes two carbon units. These enter the cycle as Acetyl-CoA 7

8 Oxidative Decarboxylation of Pyruvate - Pyruvate Dehydrogenase complex is a large multi-subunit complex located in the mitochondria - Irreversible reaction; Acetyl CoA cannot converted into pyruvate - Pyruvate dehydrogenase is not a part of citric acid cycle but it a major source of fuel for citric acid cycle which is Acetyl CoA - Pyruvate Dehydrogenase complex is aggregate of three enzymes: 1- Pyruvate dehydrogenase component called (pyruvate decarboxylase) 2- Dihydrolipoyl transacetylase 3- Dihydrolipoyl dehydrogenase Each subunit of this large complex catalyzes a part of the overall reactions. 8

9 Pyruvate Dehydrogenase: a large complex containing many copies of each of 3 enzymes, E 1, E 2, & E 3. Pyruvate Dehydrogenase Subunits Enzyme Abbreviated Prosthetic Group Pyruvate Dehydrogenase E 1 Thiamine pyrophosphate (TPP) Dihydrolipoyl Transacetylase E 2 Lipoamide 9 Dihydrolipoyl Dehydrogenase E 3 FAD

10 Cofactors for the Pyruvate Dehydrogenase include CoA-SH, NAD+, TPP, FAD, Lipolate 10

11 11

12 Regulation Pyruvate Dehydrogenase complex -Product inhibition -The enzyme complex is inhibited by Acetyl CoA when it is accumulated; the production rate is higher than the cell capacity of oxidation with citric acid cycle - High NADH/NAD+ ratio inhibits this enzyme complex Covalent modification -Two forms of the enzyme complex; - Active non- phosphorylated form - Inactive phosphorylated form -The two forms can be interconverted by the action of two enzymes phosphatase and kinase - The kinase is activated by an increase in the ratio of acetyl CoA/ CoA ratio or NADH/ NAD+. - elevated ADP\ATP ratio demand for energy inhibits the kinase and activate the phosphatase to produce more of the active non-phosphorylated 12 enzymes

13 Regulation Pyruvate Dehydrogenase (PDH) complex Pyruvate Dehydrogenase is regulated both allosterically and by reversible phosphorylation 13

14 Regulation of Pyruvate Dehydrogenase Pyruvate Dehydrogenase is regulated both allosterically and by reversible phosphorylation 14

15 15

16 16

17 4C 6C 6C 5C 4C 17

18 18

19 The Citric acid cycle 19

20 The Citric acid cycle 20

21 The Citric acid cycle 21

22 The Citric acid cycle 22

23 Control Points in the Citric Acid Cycle Citric acid cycle is controlled at two points 23

24 Control Points in the Citric Acid Cycle 24

25 Single molecule of glucose can potentially yield ~38 molecules of ATP 25

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