Carboxylase plant

Hofner, R., Vazquez-Morena, L., Winter, K., Bohnert, H.J. Schmitt, J.M. (1987). Induction of Crassulacean acid metabolism in Mesembryanthemum crystallinum by high salinity mass increase and de novo synthesis of PEP-carboxylase. Plant Physiology, 83, 915-19. 

J. F. Johnson, C. P. Vance, and D. L. Allan, Phosphorus deficiency in Lupinus aUms altered lateral root development and enhanced expression of phosphoenol-pyruvate carboxylase. Plant Physiol. 112 31 (1996). 

J.T. ChristeUer, W.A. Laing, J.H. Troughton (1976) Isotope discrimination by ribulose 1,5-diphos-phate carboxylase. Plant Physiol. 57, 580-582... 

ChoiJ.K. et al., (1995) Molecular cloning and characterization of the cDNA coding for the biotin-containing subunit of the chloroplastic acetyl-CoA carboxylase. Plant Physiol. 109 619-625... 

A. R. Slabas and A. Hellyer, Rapid purification of a high molecular weight subunit polypeptide form of rapeseed acetyl-CoA carboxylase. Plant Sci. 39 177... 

Ashton AR, Jenkins CLD,Whitfeld PR. Molecular cloning of two different cDNAs for maize acetyl-CoA carboxylase. Plant Mol Biol 1994 24 35-49. 

Egli MA, Gengenbach BG, Gronwald JW, Somers DA, Wyse DL. Characterization of maize acetyl-CoA carboxylase. Plant Physiol 1993 101 499-506. 

Figure 4.8 The active site in all a/p barrels is in a pocket formed by the loop regions that connect the carboxy ends of the p strands with the adjacent a helices, as shown schematically in (a), where only two such loops are shown, (b) A view from the top of the barrel of the active site of the enzyme RuBisCo (ribulose bisphosphate carboxylase), which is involved in CO2 fixation in plants. A substrate analog (red) binds across the barrel with the two phosphate groups, PI and P2, on opposite sides of the pocket. A number of charged side chains (blue) from different loops as welt as a Mg ion (yellow) form the substrate-binding site and provide catalytic groups. The structure of this 500 kD enzyme was determined to 2.4 A resolution in the laboratory of Carl Branden, in Uppsala, Sweden. (Adapted from an original drawing provided by Bo Furugren.)...

Pyruvate carboxylase is the most important of the anaplerotie reactions. It exists in the mitochondria of animal cells but not in plants, and it provides a direct link between glycolysis and the TCA cycle. The enzyme is tetrameric and contains covalently bound biotin and an Mg site on each subunit. (It is examined in greater detail in our discussion of gluconeogenesis in Chapter 23.) Pyruvate carboxylase has an absolute allosteric requirement for acetyl-CoA. Thus, when acetyl-CoA levels exceed the oxaloacetate supply, allosteric activation of pyruvate carboxylase by acetyl-CoA raises oxaloacetate levels, so that the excess acetyl-CoA can enter the TCA cycle. 

PEP carboxylase occurs in yeast, bacteria, and higher plants, but not in animals. The enzyme is specifically inhibited by aspartate, which is produced by transamination of oxaloacetate. Thus, organisms utilizing this enzyme control aspartate production by regulation of PEP carboxylase. Malic enzyme is found in the cytosol or mitochondria of many animal and plant ceils and is an NADPIT-dependent enzyme. 

As indicated, ribulose bisphosphate carboxylase/oxygenase catalyzes an alternative reaction in which Og replaces COg as the substrate added to RuBP (Figure 22.29a). The ribulose-l,5-bisphosphate oxygenase rezLCtion diminishes plant... 

Compartmentation of these reactions to prevent photorespiration involves the interaction of two cell types, mescrphyll cells and bundle sheath cells. The meso-phyll cells take up COg at the leaf surface, where Og is abundant, and use it to carboxylate phosphoenolpyruvate to yield OAA in a reaction catalyzed by PEP carboxylase (Figure 22.30). This four-carbon dicarboxylic acid is then either reduced to malate by an NADPH-specific malate dehydrogenase or transaminated to give aspartate in the mesophyll cells. The 4-C COg carrier (malate or aspartate) then is transported to the bundle sheath cells, where it is decarboxylated to yield COg and a 3-C product. The COg is then fixed into organic carbon by the Calvin cycle localized within the bundle sheath cells, and the 3-C product is returned to the mesophyll cells, where it is reconverted to PEP in preparation to accept another COg (Figure 22.30). Plants that use the C-4 pathway are termed C4 plants, in contrast to those plants with the conventional pathway of COg uptake (C3 plants). 

FIGURE 22.30 Essential features of the coinpartinenCation and biochemistry of die Hatch-Slack padiway of carbon dioxide uptake in C4 plants. Carbon dioxide is fixed into organic linkage by PEP carboxylase of meso-phyll cells, forming OAA. Eidier malate (die reduced form of OAA) or aspartate (the ami-iiated form) serves as die carrier transpordiig CO9 to the bundle slieadi cells. Within die bundle slieadi cells, CO9 is liberated by decar-boxyladon of malate or aspartate die C-3 product is returned to die mesophyll cell. 

Fatty acids with odd numbers of carbon atoms are rare in mammals, but fairly common in plants and marine organisms. Humans and animals whose diets include these food sources metabolize odd-carbon fatty acids via the /3-oxida-tion pathway. The final product of /3-oxidation in this case is the 3-carbon pro-pionyl-CoA instead of acetyl-CoA. Three specialized enzymes then carry out the reactions that convert propionyl-CoA to succinyl-CoA, a TCA cycle intermediate. (Because propionyl-CoA is a degradation product of methionine, valine, and isoleucine, this sequence of reactions is also important in amino acid catabolism, as we shall see in Chapter 26.) The pathway involves an initial carboxylation at the a-carbon of propionyl-CoA to produce D-methylmalonyl-CoA (Figure 24.19). The reaction is catalyzed by a biotin-dependent enzyme, propionyl-CoA carboxylase. The mechanism involves ATP-driven carboxylation of biotin at Nj, followed by nucleophilic attack by the a-carbanion of propi-onyl-CoA in a stereo-specific manner. 

CAM is unlikely to be useful for any of the conventional food crops. Nevertheless, further study of those plants in which CAM is inducible could prove useful since isolation of, for example, the PEP carboxylase gene will permit the isolation of its controlling sequences. This will provide us with another set of stress-specific (drought) promoters and enhancers. Furthermore the PEP carboxylase gene has proved amenable to cloning via the cDNA route (Harpster Taylor, 1986). 

Groenhof, A.C., Bryant, J.A. Etherington, J.R. (1988). Photosynthetic changes in the inducible CAM plant Sedum telephium L. following the imposition of water stress. II. Changes in the activity of phosphoenolpyruvate carboxylase. Annals of Botany, 62, 187-92. 

Ostrem, J.A., Olson, S.W., Schmitt, J.M. Bohnert, H.J. (1987). Salt stress increases the level of translatable mRNA for phosphoenolpyruvate carboxylase in Mesembryanthemum crystallinum. Plant Physiology, 84,1270-5. 

An intriguing stress-induced alteration in gene expression occurs in a succulent plant, Mesembryanthemum crystallinum, which switches its primary photosynthetic CO2 fixation pathway from C3 type to CAM (Crassulacean acid metabolism) type upon salt or drought stress (Winter, 1974 Chapter 8). Ostrem et al. (1987) have shown that the pathway switching involves an increase in the level of mRNA encoding phosphoenol-pyruvate carboxylase, a key enzyme in CAM photosynthesis. 

Mayoral, M.L., Atsmon, D., Gromet-Elhanan, Z. Shimshi, D. (1981). Effect of water stress on enzyme activities in wheat and related wild species Carboxylase activity, electron transport and photophosphorylation in isolated chloroplasts. Australian Jourrml of Plant Physiology, 8, 385-94. 

Vierling, E. Key, J.L. (1985). Ribulose 1,5-bisphosphate carboxylase synthesis during heat shock. Plant Physiology, 78,155-62. 

Nonphotosynthetic COi fixation via phosphoenolpyruvate carboxylase (PEPC) can contribute a substantial proportion of carbon (>30%) for the biosynthesis of carboxylates in roots of P-deficient plants (Fig. 5) (11,82,101,111-113). Thus, PEPC-mediated COi fixation may be interpreted as an anaplerotic carbon... 

Atmospheric CO2 first moves through the stomata, dissolves into leaf water and enters the outer layer of photosynthetic cells, the mesophyll cell. Mesophyll CO2 is directly converted by the enzyme ribulose biphosphate carboxylase/oxygenase ( Rubisco ) to a six carbon molecule that is then cleaved into two molecules of phosphoglycerate (PGA), each with three carbon atoms (plants using this photosynthetic pathway are therefore called C3 plants). Most PGA is recycled to make ribulose biphosphate, but some is used to make carbohydrates. Free exchange between external and mesophyll CO2 makes the carbon fixation process less efficient, which causes the observed large C-depletions of C3 plants. 

C4 plants incorporate CO2 by the carboxylation of phosphoenolpyruvate (PEP) via the enzyme PEP carboxylase to make the molecule oxaloacetate which has 4 carbon atoms (hence C4). The carboxylation product is transported from the outer layer of mesophyll cells to the inner layer of bundle sheath cells, which are able to concentrate CO2, so that most of the CO2 is fixed with relatively little carbon fractionation. 

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