Abstract
In order to determine whether the enzymes required to convert triose phosphate to acetyl CoA were present in pea (Pisum sativum L.) seed plastids, a rapid, mechanical technique was used to isolate plastids from developing cotyledons. The plastids were intact and the extraplastidial contamination was low. The following glycolytic enzymes, though predominantly cytosolic, were found to be present in plastids: glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12), phosphoglycerate kinase (EC 2.7.2.3), and pyruvate kinase(EC 2.7.1.40). Evidence is presented which indicates that plastids also contained low activities of enolase (EC 4.2.1.11) and phosphoglycerate mutase (EC 2.7.5.3). Pyruvate dehydrogenase, although predominantly mitochondrial, was also present in plastids. The plastidial activities of the above enzymes were high enough to account for the rate of lipid synthesis observed in vivo.
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Abbreviations
- FPLC:
-
fast protein liquid chromatography
- PPi:
-
pyrophosphate
References
ap Rees, T., Bryce, J.H., Wilson, P.M.,Green, J.H. (1983) Role and location of NAD malic enzyme in the thermogenic tissues of Araceae. Arch. Biochem. Biophys. 227, 511–521
ap Rees, T., Thomas, S.M., Fuller, W.A., Chapman, B. (1975) Location of gluconeogenesis from phosphoenolpyruvate in cotyledons of Cucurbita pepo. Biochim. Biophys. Acta. 385, 145–156
Brooks, J.L., Stumpf, P.K.(1966) Fat metabolism in higher plants. XXXIX. Properties of a soluble fatty acid synthesising system from lettuce chloroplasts. Arch. Biochem. Biophys. 166, 108–116
Camp, P.J., Randall, D.D. (1985) Purification and characterization of pea chloroplast pyruvate dehydrogenase complex. Plant Physiol. 77, 571–577
Coxon, D.T., Davies, D.R. (1982) The effect of the r a and r b loci on the lipid content of the seed of Pisum sativum. Theor. Appl. Genet. 64, 47–50
Elias, B.A., Givan, C.V. (1979) Localization of pyruvate dehydrogenase complex in Pisum sativum chloroplasts. Plant Sci. Lett. 17, 115–122
Flügge, U.I., Heldt, H.W. (1984) The phosphate-triose phosphate-phosphoglycerate translocator of the chloroplast. Trends Biochem. Sci. 108, 530–533
Gerhardt, R., Heldt, H.W.(1984) Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in non-aqueous media. Plant Physiol. 75, 542–547
Hedley, C.L., Ambrose, M.J., Smith, C.M., Cook, S., Wang, T.L. (1986a) Redesigning the pea seed. In: Proc. EUCAR-PIA Meet on Pea Breeding, pp. 147–163, Monti, L., ed. Sorrento, University of Naples
Hedley, C.L., Smith, C.M., Ambrose, M.J., Cook, S., Wang, T.L. (1986b) An analysis of seed development in Pisum sativum. II. The effect of the r-locus on the growth and development of the seed. Ann. Bot. 58, 371–379
Hodges, T.K., Leonard, R.T. (1974) Purification of a plasma membrane bound adenosine triphosphatase from plant roots. Methods Enzymol. 32, 392–406
Ireland, R.J., DeLuca, V., Dennis, D.T. (1980) Characterization and kinetics of isoenzymes of pyruvate kinase from developing castor bean endosperm. Plant Physiol. 65, 1188–1193
Journet, E., Douce, R. (1985) Enzymic capacities of purified cauliflower bud plastids for lipid synthesis and carbohydrate metabolism. Plant Physiol. 79, 458–467
Kuhn, D.N., Knauf, M., Stumpf P.K. (1981) Subcellular localization of acetyl CoA synthetase in leaf protoplasts of Spinacia oleracea. Arch. Biochem. Biophys. 209, 441–450
Liedvogel, B., Bäuerle, R. (1986) Fatty-acid synthesis in chloroplasts from mustard (Sinapis alba L.) cotyledons: formation of acetyl coenzyme A by intraplastid glycolytic enzymes and a pyruvate dehydrogenase complex. Planta 169, 481–489
Liedvogel, B., Stumpf, P.K. (1982) Origin of acetate in spinach leaf cell. Plant Physiol. 69, 897–903
MacDonald, F.D., ap Rees, T. (1983) Enzymic properties of amyloplasts from suspension cultures of soybean. Biochim. Biophys. Acta. 755, 81–89
Miernyk, J.A., Dennis, D.T. (1982) Isoenzymes of the glycolytic enzymes in endosperm from developing castor oil seeds. Plant Physiol. 69, 825–828
Proudlove, M.D., Thurman, D.A. (1981) The uptake of 2-oxoglutarate and pyruvate by isolated pea chloroplasts. New Phytol. 88, 255–264
Reynolds, E.S. (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–212
Rubin, P.M., Randall, D.D. (1977) Purification and characterization of pyruvate dehydrogenase complex from broccoli floral buds. Arch. Biochem. Biophys. 178,342–349
Scopes, R.K. (1975) 3-phosphoglycerate kinase of skeletal muscle. Methods Enzymol. 42, 127–134
Simcox, D.A., Reid, E.E., Canvin, D.T., Dennis, D.T. (1977) Enzymes of the glycolytic and pentose phosphate pathways in proplastids from the developing endosperm of Ricinus communis L. Plant. Physiol. 59, 1128–1132
Smith, A.M., ap Rees, T. (1979) Effects of anaerobiosis on carbohydrate oxidation by roots of Pisum sativum. Phytochemistry 18, 1453–1458
Stitt, M., ap Rees, T. (1979) Capacities of pea chloroplasts to catalyse the oxidative pentose phosphate pathway and glycolysis. Phytochemistry 18, 1905–1911
Thorne, J.H. (1985) Phloem unloading of C and N assimilates in developing seeds. Annu. Rev. Plant Physiol. 36, 317–343
Tolbert, N.E. (1974) Isolation of subcellular organelles of metabolism on isopycnic sucrose gradients. Methods Enzymol. 31, 734–747
Turner, J.F. (1969) Physiology of pea fruits. IV. Changes in uridine diphosphate glucose pyrophosphorylase and adenosine diphosphate glucose pyrophosphorylase in the developing seed. Aust. J. Biol. Sci. 22, 1145–1151
Wong, K.F., Davies, D.D. (1973) Regulation of phosphoenolpyruvate carboxylase of Zea mays by metabolites. Biochem. J. 131, 451–458
Wu, R., Racker, E. (1959) Regulatory mechanisms in carbohydrate metabolism. III. Limiting factors in glycolysis of ascites tumor cells. J. Biol. Chem. 234, 1029–1035
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Denyer, K., Smith, A.M. The capacity of plastids from developing pea cotyledons to synthesise acetyl CoA. Planta 173, 172–182 (1988). https://doi.org/10.1007/BF00403008
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DOI: https://doi.org/10.1007/BF00403008