Citrate cycle

Two and twelve moles of ATP are produced, respectively, per mole of glucose consumed in the glycolytic pathway and each turn of the Krebs (citrate) cycle. In fat metaboHsm, many high energy bonds are produced per mole of fatty ester oxidized. Eor example, 129 high energy phosphate bonds are produced per mole of palmitate. Oxidative phosphorylation has a remarkable 75% efficiency. Three moles of ATP are utilized per transfer of two electrons, compared to the theoretical four. The process occurs via a series of reactions involving flavoproteins, quinones such as coenzyme Q, and cytochromes. 

If the glycolytic flux is slow much of the pyruvate formed enters the mitochondria and is oxidized by the citrate cycle and reducing equivalents (2H) from NADH are oxidized indirectly (see below). When the flux is fast there is net production of... 

Figure 3. Mitochondrial fatty acid oxidation. Long-chain fatty acids are converted to their CoA-esters as described in the text, and their fatty-acyl-groups transferred to CoA in the matrix by the concerted action of CPT 1, the acylcarnitine/carnitine exchange carrier and CPT (A) as described in the text. Medium-chain and short-chain fatty acids (Cg or less) diffuse directly into the matrix where they are converted to their acyl-CoA esters by a acyl-CoA synthase. The mechanism of p-oxidation is shown below (B). Each cycle of P-oxidation removes -CH2-CH2- as an acetyl unit until the fatty acids are completely converted to acetyl-CoA. The enzymes catalyzing each stage of P-oxidation have different but overlapping specificities. In muscle mitochondria, most acetyl-CoA is oxidized to CO2 and H2O by the citrate cycle (Figure 4) some is converted to acylcamitine by carnitine acetyltransferase (associated with the inner face of the inner membrane) and exported from the matrix. Some acetyl-CoA (if in excess) is hydrolyzed to acetate and CoASH by acetyl-CoA hydrolase in the matrix. Enzymes ...

In peripheral tissues acetoacetate exported by the liver reacts with succinyl-CoA formed in the citrate cycle to give acetoacetyl-CoA and succinate catalyzed by a specific CoA transferase. 

CH3COCH2COSC0A + CoASH which are then oxidized by the citrate cycle. 

The metabolism of amino acids is complex and is described in standard text books. These are usually converted by aminotransferases to the corresponding 2-oxoacids which are partly oxidized in the matrix of muscle mitochondria and partly exported to the liver. Glutamate and aspartate yield 2-oxoglutarate and oxaloacetate, respectively, which enter the citrate cycle directly, and other 2-... 

The citrate cycle is the final common pathway for the oxidation of acetyl-CoA derived from the metabolism of pyruvate, fatty acids, ketone bodies, and amino acids (Krebs, 1943 Greville, 1968). This is sometimes known as the Krebs or tricarboxylic acid cycle. Acetyl-CoA combines with oxaloacetate to form citrate which then undergoes a series of reactions involving the loss of two molecules of CO2 and four dehydrogenation steps. These reactions complete the cycle by regenerating oxaloacetate which can react with another molecule of acetyl-CoA (Figure 4). 

It has often been questioned whether the rates and kinetics of purified enzymes, determined in very dilute solutions with high concentrations of their substrates, but not always of their cofactors, can be extrapolated to the conditions prevailing in the matrix. Much of the mitochondrial water will be bound to protein by hydrogen bonds and electrostatically, but there is also a pool of free water which may only be a fraction of the total water (Gitomer, 1987). The molar concentrations of intermediates of the citrate cycle and of p-oxidation are very low, usually less than those of most enzymes (Srere, 1987 Watmough et al., 1989 Sumegi et al., 1991). The extent to which cofactors and intermediates bind specifically or nonspecifically to enzymes is not known. It is therefore difficult to estimate concentration of these... 

Figure 4. The citrate cycle. There is complete oxidation of one molecule of acetyl-CoA for each turn of the cycle CH3COSC0A + 2O2 - 2CO2 + H2O + CoASH. The rate of the citrate cycle is determined by many factors including the ADP/ATP ratio, NAD7NADH ratio, and substrate concentrations. During muscle contraction, Ca is released from cellular stores (mainly the sarcoplasmic reticulum) and then taken up in part by the mitochondria (see Table 2). Ca " activates 2-oxoglutarate and isocitrate dehydrogenases (Brown, 1992). Succinate dehydrogenase may be effectively irreversible. Enzymes ...

Wachtcrshauser s prime candidate for a carbon-fixing process driven by pyrite formation is the reductive citrate cycle (RCC) mentioned above. Expressed simply, the RCC is the reversal of the normal Krebs cycle (tricarboxylic acid cycle TCA cycle), which is referred to as the turntable of metabolism because of its vital importance for metabolism in living cells. The Krebs cycle, in simplified form, can be summarized as follows ... 

The formation of acetate CH3C02 + H+ from C02 and CH4. The acetyl group CH3CO- is the original building block of other carboxylic acids, by the reverse citrate cycle (Figure 4.4), and of lipids in cells. 

The addition of ammonia to the variety of acids derivable from either the breakdown of glucose, glycolysis, or of the pentose shunt reaction products, ribose and NADPH, and from the citrate cycle, gives the amino acids (see Table 4.7 and Figure 4.4) Polymerisation of amino acids in cells gives proteins. In some of the amino acids sulfur and selenium can be incorporated easily. We assume NH3 was present. (Note that Se is in a coded amino acid not in Table 4.7.) Some selective metal-binding properties can be seen in Table 4.7, but amino acid carboxylates can bind all. 

Fig. 9. Pathway duplication the methyl citrate cycle and the glyoxylate shunt. A pathway for acetate metabolism in E. coli that uses the glyoxylate shunt is depicted on the right. Part of the methyl citrate cycle, a pathway for propionate metabolism, is depicted on the left. The pathways are analogous furthermore, three of the four steps are catalyzed by homologous enzymes. PrpE (propionyl-CoA synthase) is homologous to AcsA (acetyl-CoA synthase). PrpC (2-methyl-citrate synthase) is homologous to GltA (citrate synthase). PrpB (2-methyl-isocitrate lyase) is homologous to AceA (isocitrate lyase). The third step in the methyl citrate cycle has been suggested to be catalyzed by PrpD the second half of the reaction (the hydration) can be catalyzed by aconitase.

Carbohydrate Metabolism Glycolysis/ Gluconeogenesis Citrate cycle (TCA cycle)... 

When the oxidation of substantial amounts of pyruvate was blocked by malonate, stoichiometric amounts of pyruvate would react if either oxaloacetate or its precursors, malate or fumarate, was added. None of the precursors to succinate in the chain was effective in this regard. This strongly supported Krebs hypothesis that oxidation of pyruvate involved its condensation with oxaloacetate and the subsequent series of conversions in the chain described by Krebs. The Krebs cycle in its originally proposed form is shown in figure 13.3. It is also known as the tricarboxylic acid (TCA) cycle or the citrate cycle. 

Original tricarboxylic acid (TCA) cycle proposed by Krebs. This cycle is also called the citrate cycle, or the Krebs cycle. To start the cycle in operation, pyruvate loses one of its carbons and condenses with a four-carbon dicarboxylic acid, oxaloacetic acid, to form a six-carbon tricarboxylic acid, citrate. In one turning of the cycle, two carbons are lost as C02, thus returning the citrate to oxaloacetate. The conversion blocked by malonate is indicated by a red bar. 

Eu(Tc)(cit) is 1 1 2 in this case. The fluorescence intensity of the 615-nm emission line of [Eu(Tc)(cit)2 ] is 22 times stronger than that of [Eu(Tc)]. Citrate, as a polydentate ligand, can chelate the Eu3+ ion via the oxygen atoms of its carboxy and hydroxy groups. It is assumed that citrate displaces water molecules which ligate to the 8- and/or 9-coordination sites of the Eu3+ ion and quench its fluorescence. Table 5 summarizes the luminescence properties of the [Eu(Tc)] complex (1 1 stoichiometry) and the corresponding chelate complexes with the intermediates of the citrate cycle. Citrate concentrations can be imaged with this luminescent probe by means of the RLI method (Fig. 15) [107]. 

Lin Z (2004) Time-resolved fluorescence-based europium-derived probes for peroxidase bioassays, citrate cycle imaging and chirality sensing. PhD thesis, University of Regensburg... 

Knappe, E. and Rohdewald, I., Thin-layer chromatography of dicarboxylic acids. V. Separation and identification of hydroxy dicarboxylic acids, of di- and tricarboxylic acids of the citrate cycle, and some other dicarboxylic acids of plant origin, Z. Anal. Chem., 211, 49, 1965 Chem. Abs., 63, 7333c, 1965. 

To give a real example, have a closer look on main functions and cycle of magnesium in green plants. Control on autocatalysis depends on the principal functions of Mg, that is, on photosynthesis when substantial parts of Mg taken up by roots are allocated to chlorophyll and rubisco synthesis, less will be available for other metabolic pathways, reducing the turnovers there unless there are lots of Mg around like in marine plants. In addition, the tricarboxylate cycle (citrate cycle) requires Mg (besides Fe and Mn) to produce the enzymes hence some Mg (as well as Fe, Mn) must be invested to produce the citrate (malate, oxaloacetate (aspartate)) ions delivered by the roots to render Mg (and other metals) in turn bioavailable by means of complexation and resorption of almost neutral complex entities. Furthermore, the tricarboxylate cycle is coupled to biosynthesis of amino acids by redox transamination hence there will be both competition at the metal center(s) and possible extraction of metal ions from enzymes once NHj and electrons are... 

Amino acid synthesis from citrate cycle products which indirectly extract Mg from the citrate cycle because the corresponding complexes are more stable than those with citrate, malate. .., reducing... 

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