Krebs Cycle | Research & Encyclopedia Articles

Krebs Cycle

The Krebs cycle is a set of biochemical reactions that occur in the mitochondria. The Krebs cycle is the final common pathway for the oxidation of food molecules such as sugars and fatty acids. It is also the source of intermediates in biosynthetic pathways, providing carbon skeletons for the synthesis of amino acids, nucleotides, and other key molecules in the cell. The Krebs cycle is also known as the citric acid cycle, and the tricarboxylic acid cycle. The Krebs cycle is a cycle because, during its course, it regenerates one of its key reactants.

To enter the Krebs cycle, a food molecule must first be broken into two- carbon fragments known as acetyl groups, which are then joined to the carrier molecule coenzyme A (the A stands for acetylation). Coenzyme A is composed of the RNA nucleotide adenine diphosphate, linked to a pantothenate, linked to a mercaptoethylamine unit, with a terminal S-H.Dehydration of this linkage with the OH of an acetate group produces acetyl CoA. This reaction is catalyzed by pyruvate dehydrogenase complex, a large multi-enzyme complex.

The acetyl CoA linkage is weak, and it is easily and irreversibly hydrolyzed when Acetyl CoA reacts with the four-carbon compound oxaloacetate. Oxaloacetate plus the acetyl group form the six-carbon citric acid, or citrate. (Citric acid contains three carboxylic acid groups, hence the alternate names for the Krebs cycle.)

Following this initiating reaction, the citric acid undergoes a series of transformations. These result in the formation of 3 molecules of the high-energy hydrogen carrier NADH (nicotinamide adenine dinucleotide), 1 molecule of another hydrogen carrier FADH₂ (flavin adenine dinucleotide), 1 molecule of high-energy GTP (guanine triphosphate), and 2 molecules of carbon dioxide, a waste product. The oxaloacetate is regenerated, and the cycle is ready to begin again. NADH and FADH₂ are used in the final stages of cellular respiration to generate large amounts of ATP.

The details of the cycle are as follows:

1. Oxaloacetate condenses with acetyl coA to form citrate. This reaction is catalyzed by the enzyme citrate synthase.

2. Citrate undergoes isomerization to isocitrate, via a dehydration/rehydration step involving the temporary creation of cis-aconitate. This reaction, catalyzed by aconitase, prepares the molecule for decarboxylation in the next step.

3. Isocitrate undergoes an oxidative decarboxylation, in which it both loses a CO₂ molecule and becomes more oxidized. Isocitrate is first converted to oxalosuccinate by removal of a hydrogen, and formation of NADH. Oxalosuccinate is then decarboxylated, becoming the five-carbon compound alpha-ketoglutarate. This reaction is catalyzed by isocitrate dehydrogenase.

4. Alpha-ketoglutarate is itself decarboxylated, forming another NADH and CO₂. The four-carbon product, succinate, is temporarily linked to coenzyme A, forming the weakly linked succinyl CoA. This reaction is catalyzed by the alpha- ketoglutarate dehydrogenase complex.

5. Succinyl CoA is cleaved by succinyl CoA synthetase to release succinate. In the process, guanosine diphosphate (GDP) is linked to inorganic phosphate to form guanosine triphosphate (GTP). GTP is used in the cell for protein synthesis and signal cascades, or the energy can be transferred to create ATP for most common energy-requiring tasks.

6. The four-carbon succinate undergoes a series of rearrangements to regenerate oxaloacetate. It is first oxidized to form fumarate (catalyzed by succinate dehydrogenase), with a hydrogen transfer to FAD to form FADH₂. Fumarate is hydrated by fumarase to form malate, which is then oxidized by malate dehydrogenase to form oxaloacetate, with the creation of a final NADH.

As a central metabolic pathway in the cell, the rate of the Krebs cycle must be tightly controlled to prevent too much, or too little, formation of products. This regulation occurs through inhibition or activation of several of the enzymes involved. Most notably, the activity of pyruvate dehydrogenase is inhibited by its products, acetyl CoA and NADH, as well as by GTP. This enzyme can also be inhibited by enzymatic addition of a phosphate group, which occurs more readily when ATP levels are high. Each of these actions serves to slow down the Krebs cycle when energy levels are high in the cell. it is important to note that the Krebs cycle is also halted when the cell is low on oxygen, even though no oxygen is consumed in it. Oxygen is needed further along in cell respiration though, to regenerate NAD⁺ and FAD. Without these, the cycle cannot continue, and pyruvic acid is converted in the cytosol to lactic acid by the fermentation pathway.

The Krebs cycle is also a source for precursors for biosynthesis of a number of cell molecules. For instance, the synthetic pathway for amino acids can begin with either oxaloacetate or alpha-ketoglutarate, while the production of porphyrins, used in hemoglobin and other proteins, begins with succinyl CoA. Molecules withdrawn from the cycle for biosynthesis must be replenished. Oxaloacetate, for instance, can be formed from pyruvate, carbon dioxide, and water, with the use of one ATP molecule.