CITRIC ACID CYCLE

DR. M ALI MASOOD LECTURER DCOP,DUHS

CITRIC ACID CYCLE

The citric acid cycle (krebs cycle or tricarboxylic acid-TCA cycle) is the most important metabolic pathway for the energy supply to the body. About 65-70% of the ATP is synthesized in krebs cycle. Citric acid cycle essentially involves the oxidation of acetyl CoA to CO2 and H2O. This cycle utilizes about two-thirds of total oxygen consumed by the body. The name TCA cycle is used, since, at the outset of the cycle, tricarboxylic acids (citrate , cis-aconitate and isocitrate) participate.

TCA cycle-the central metabolic pathway

The citric acid cycle is the final common oxidative pathway for carbohydrates, fats and amino acids. This cycle not only supplies energy but also provides many intermediates required for the synthesis of amino acids, glucose , heme etc. Krebs cycle is the most important central pathway connecting almost all the individual metabolic pathways (either directly or indirectly).

BRIEF HISTORY
The citric acid cycle was proposed by Hans Adolf Krebs in 1937, based on the studies of oxygen consumption in pigeon breast muscle. The cycle is named in his honour (Nobel Prize for physiology and Medicine in 1953). LOCATION OF TCA-CYCLE The enzymes of TCA cycle are located in mitochondrial matrix. This enables the synthesis of ATP by oxidative phosphorylation without any hindrance.

TCA CYCLE AN OVERVIEW

Krebs cycle basically involves the combination of a two carbon acetyl CoA with a four carbon oxaloacetate to produce a six carbon tricarboxylic acid, citrate. In the reactions that follow, the two carbons are oxidized to CO2 and oxaloacetate is regenerated and recycled. Oxaloacetate is considered to play a catalytic role in citric acid cycle.

Citric Acid Cycle

TCA cycle an open cycle.

Krebs cycle is a cyclic process. However, it should not be viewed as a closed circle, since many compounds enter the cycle and leave. TCA cycle is comparable to a heavy traffic circle in a national highway with many connecting roads. Each intermediate of the cycle connecting another pathway is a road.

REACTIONS OF CITRIC ACID CYCLE

Oxidative decarboxylation of pyruvate to acetyl CoA by pyruvate dehydrogenase complex is previously discussed . This step is a connecting link between glycolysis and TCA cycle.
1. Formation of citrate: Krebs cycle proper starts with the condensation of acetyl CoA and oxaloacetate, catalysed by the enzyme citrate synthase.

Citrate Cycle: step 1 (citrate formation)

Enzyme: Citrate synthase

Condensation reaction

Hydrolysis reaction

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2 & 3. Citrate is isomerized to isocitrate by the enzyme aconitase. This is achieved in a two stage reaction of dehydration followed by hydration through the formation of an intermediate cis-aconitate.

Citrate isomerized to Isocitate: step 2 & 3

Enzyme: aconitase

Dehydration

Hydration

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4. & 5. Formation of -Ketoglutarate: The enzyme isocitrate dehydrogenase (ICD) catalyses the conversion (oxidative decarboxylation) of isocitrate to oxalosuccinate and then to -Ketoglutarate. The formation of NADH and the liberation of CO2 occur at this stage.

Isocitrate to -ketoglutarate: step 4 & 5

Enzyme: isocitrate dehydrogenase

1st NADH produced

1st CO2 removed

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6. Conversion of -ketoglutarate to succinyl CoA occur through oxidative decarboxylation, catalysed by ketoglutarate dehydrogenase complex. This enzyme is dependent on five cofactorsTPP, lipoamide, NAD+, FAD and CoA. The mechanism of the reaction is analogous to the conversion of pyruvate to acetyl CoA. At this stage of the TCA cycle, second NADH is produced and the second Co2 is liberated.

Succinyl CoA formation: step 6

Enzyme: -ketoglutarate dehydrogenase

2nd NADH produced

2nd CO2 removed

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7. Formation of succinate: Succinyl CoA is converted to succinate by succinyl CoA synthetase. This reaction is coupled with the phosphorylation of GDP to GTP. This is a substrate level phosphorylation. GTP is converted to ATP by the enzyme nucleoside diphosphate kinase.
GTP + ADP ATP + GDP

Succinate formation: step 7

Enzyme: succinyl CoA synthetase

GTP produced GTP + ADP GDP + ATP (NPTase)

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8. Conversion of succinate to fumarate: Succinate is oxidized by succinate dehydrogenase to fumarate. This reaction result in the production of FADH2 & not NADH.

Succinate to Fumarate: step 8

Enzyme: succinate dehydrogenase

FADH2 produced

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9. Conversion of malate: The enzyme fumarase catalysis the conversion of fumarate to malate with the addition of water.

Fumarate to Malate: step 9

Enzyme: fumarase

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10. Conversion of malate to oxaloacetate: Malate is then oxidized to oxaloacetate by malate dehydrogenase. The third and final synthesis of NADH occurs at this stage. The oxaloacetate is regenerated which can combine with another molecule of acetyl CoA, and continue the cycle

Malate to Oxalate: step 10

Enzyme: malate dehydrogenase

3rd NADH produced

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Summary of TCA cycle

The events of Krebs cycle may be summarized as given in the next column.
Acetyl CoA + 3NAD+ + FAD + GDP +Pi +2H2O 3NADH + 3H+ + FADH2 + GTP + CoA 2CO2 +

Proton gradient generates 3ATP per NADH, & 2 per FADH2 9 ATP from 3 NADH + 2 from FADH2. Also, 1 from GTP Thus, 1 acetate unit generates equivalent of 12 ATP molecules.

Energy Produced By TCA Cycle

Four pairs of electrons are transferred during one turn of the cycle: three pairs of electrons reducing NAD+ to NADH and one pair reducing FAD to FADH2. Oxidation of one NADH by the electron transport chain leads to formation of approximately three ATP, whereas oxidation of FADH2 yields approximately two ATP.

Regeneration of oxaloacetate in TCA cycle

The TCA cycle basically involves the oxidation of acetyl CoA to Co2 with simultaneouly regeneration of oxaloacetate. As such, there is no net consumption of oxaloacetate or any other intermediate in the cycle.

REQUIREMENT OF O2 BY TCA CYCLE

There is no direct participation of oxygen in Krebs

cycle. However, the cycle operates only under aerobic conditions.

AMPHIBOLIC NATURE OF THE CITRIC ACID CYCLE.

The citric acid cycle provides various intermediates for the synthesis of many compounds needed by the body. Krebs cycle is both catabolic and anabolic in nature, hence regarded as amphibolic. TCA cycle is actively involved in gluconeogenesis, transamination and deamination.

1. Oxaloacetate and ketoglutarate, respectively, serve as precursors for the synthesis of aspartate and glutamate which, in turn, are required for the synthesis of other non-essential amino acids, purines and pyrimidines. 2. Succinyl CoA is used for the synthesis of porphyrins and heme. 3. Mitochondrial citrate is transported to the cytosol, where it is cleaved to provide acetyl CoA for the biosynthesis of fatty acids, sterols etc.

Biosynthetic roles of the citric acid cycle

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