Tricarboxylic acid cycle control

Coenzyme availability can also often have a limiting effect (5). If the coenzyme is regenerated by a second, independent metabolic pathway, the speed of the second pathway can limit that of the first one. For example, glycolysis and the tricarboxylic acid cycle are mainly regulated by the availability of NAD" (see p. 146). Since NAD is regenerated by the respiratory chain, the latter indirectly controls the breakdown of glucose and fatty acids (respiratory control, see p. 144). 

All plants contain a PEPCase enzyme which, among other likely roles (Vidal et al., 1986), serves to replenish tricarboxylic acid cycle intermediates that are consumed during ammonium assimilation (Latzko Kelly, 1983). The role of this enzyme, other than its presumed housekeeping function, has not been studied in any detail and its activity is probably not controlled by environmental factors. Isoforms of PEPCase have been observed in many plants (C3, C4 and CAM) for example, in rice five isoforms have been detected immunologically and in most plants two to four bands that react with anti-PEPCase antibodies are found (Matsuoka Hata, 1987). It is not clear how these isoforms arise, e.g. by post-translational modification of one form, or whether all of these forms are products of different genes. The housekeeping-type PEPCase enzyme, concerned with anaplerotic functions, is distinct from other PEPCase enzymes that function in plants with C4 and CAM metabolism (see below). 

Figure 10-1. Enzymatic pathways for glucose synthesis from amino acids or pyruvate in mammalian Ever. Enclosed in the boxes are the glucogenic amino acids with arrows indicating the points where carbon skeletons from these amino acids enter the pathways of gluconeogenesis or the tricarboxylic acid cycle. Bracketed next to the rate-controlling enzymes for gluconeogenesis are some of the substances that increase (T) or decrease (1) the activity of these enzymes. 3PG, 3-phosphoglycerate.

How could the activities of the kinase and phosphorylase be regulated so as to control the entry of pyruvate into the tricarboxylic acid cycle ... 

Much has been published on the controversial subject of the control of glycolysis. The following brief summary of some of the controls responsible for the Pasteur effect in yeasts is based mainly on a review by Sols and coworkers144 (see also, Fig. 7). (i) Isocitrate dehydrogenase (NAD ) (EC 1.1.1.41), one of the controlling enzymes of the tricarboxylic acid cycle (see Fig. 5), catalyzes the reaction... 

When pyruvate- C is injected into an injured rat, the label does not pass downward to CO2 and the amino acids glutamate and aspartate associated with the tricarboxylic acid cycle, as it does in the controls, and a greater proportion of it migrates upward toward glucose. In consequence, there is a greater labeling of the blood glucose of the injured... 

The citric acid cycle, also known as the tricarboxylic acid cycle or the Krebs cycle, is the final oxidative pathway for carbohydrates, lipids, and amino acids. It is also a source of precursors for biosynthesis. The authors begin Chapter 17 with a detailed discussion of the reaction mechanisms of the pyruvate dehydrogenase complex, followed by a description of the reactions of the citric acid cycle. This description includes details of mechanism and stereospecificity of some of the reactions, and homologies of the enzymes to other proteins. In the following sections, they describe the stoichiometry of the pathway including the energy yield (ATP and GTP) and then describe control mechanisms. They conclude the chapter with a summary of the biosynthetic roles of the citric acid cycle and its relationship to the glyoxylate cycle found in bacteria and plants. 

They catalyse rate-limiting steps in the pathway and are important control points. Glucogenic intermediates of the tricarboxylic acid cycle and amino acids that are transaminated or deaminated to tricarboxylic acid intermediates do not require pyruvate carboxylase (Fig. 3.1). Pyruvate and metabolites such as lactate, alanine, serine, glycine, cysteine that are converted into pyruvate require all four enzymes (Fig. 3.1). Gluconeogenesis is often linked to glycogen synthesis catalysed by glycogen synthase (see Section 3.5). 

The metabolic steps in gluconeogenesis occur in two intracellular compartments (Fig. 3.2) the cytosol and the mitochondrial matrix. The enzymes of the tricarboxylic acid cycle reside in the mitochondrial matrix, apart from succinate dehydrogenase which is present in the inner mitochondrial membrane, whereas most of the enzymes of the gluconeogenic pathway are present in the cytosol. Transaminases, such as alanine aminotransferase and aspartate aminotransferase, are present both in mitochondria and cytosol of the domestic fowl liver (Sarkar, 1977). One of the control enzymes in gluconeogenesis, PEPCK, has a different intracellular distribution in avian liver compared with mammalian liver (Table 3.3). PEPCK in both pigeon and domestic fowl liver is present almost exclusively (> 99%) in mitochondria (Soling et al.. 1973), whereas in most mammals that have been studied, it is present mainly in the cytosol, and only present, if at all, in smaller amounts in... 

Drynan, L., Quant, P.A. Zammit, V.A. ( 996) Biochem. J. 317, 791-795. Flux control exerted by mitochondrial outer membrane carnitine palmitoyltransferase over P-oxidation, ketogenesis and tricarboxylic acid cycle activity in hepatocytes isolated from rats in different metabolic states. 

Tricarboxylic Acid Cycle Intermediates and the Control of Fatty Acid Synthesis and Ketogenesis... 

The effect of different amino acids supplements on the synthesis of PHB by recombinant E. coli was evaluated by Mahishi and Rawal. The study revealed that when the basal medium is supplemented with amino acids, except glycine and valine, all other amino acid supplements enhanced PHB accumulation in recombinant E. coli harboring PHB synthesizing genes from S. aureqfaciens. Cysteine, isoleucine, or methionine supplementation increased PHB accumulation by 60, 45, and 61%, respectively. Amino acid biosynthetic enzyme activities in several pathways are repressed by end produa supplementation. End product inhibition in the cysteine biosynthetic pathway controls the carbon flow due to sensitivity of serine transacetylase to cysteine. Hence, supplementation of cysteine favors a change in carbon flux that eliminates the requirement of acetyl-CoA for serine transacetylation which in turn provides more carbon source and acetyl-CoA for PHB synthesis. Degradation of methionine and isoleucine yields succinyl CoA, an intermediate of tricarboxylic acid cycle and allows more acetyl-CoA to enter the PHB biosynthetic pathway. 

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