Abstract
Soluble starch synthase was purified 10000-fold from developing embryos of pea (Pisum sativum L.). The activity was resolved into two forms which together account for most if not all of the soluble starchsynthase activity in the embryo. The two isoforms differ in their molecular weights but are similar in many other respects. Their kinetic properties are similar, neither isoform is active in the absence of primer, and both are unstable at high temperatures, the activity being abolished by a 20-min incubation at 45° C. Both isoforms are recognised by antibodies raised to the granule-bound starch synthase of pea. Isoform II, which has the same molecular weight (77 kDa) as the granulebound enzyme, is recognised more strongly than Isoform I.
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Abbreviations
- DEAE:
-
diethylaminoethyl
- Hepes:
-
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Baba, T., Noro, M., Hiroto, M., Arai, Y. (1990) Properties of primer-dependent starch synthesis catalysed by starch synthase from potato tubers. Phytochemistry 29, 719–723
Bhattacharyya, M.K., Smith, A.M., Ellis, T.H.N., Hedley, C., Martin, C. (1990) The wrinkled-seed character of pea described by Mendel is caused by a transposon-like insertion in a gene encoding starch-branching enzyme. Cell 60, 115–121
Boyer, C. (1985) Soluble starch synthases and starch branching enzymes from developing seeds of sorghum. Phytochemistry 24, 15–18
Caley, C.Y., Duffus, C.M., Jeffcoat, B. (1990) Effects of elevated temperature and reduced water uptake on enzymes of starch synthesis in developing wheat grains. Aust. J. Plant Physiol. 17, 431–439
Chowdhury, S.I., Wardlaw, I.F. (1978) The effect of temperature on kernel development in cereals. Aust. J. Agric. Res. 29, 205–223
Dang, P.L., Boyer, C.D. (1988) Maize leaf and kernel starch synthases and starch branching enzymes. Phytochemistry 27, 1255–1259
Denyer, K., Smith, A.M. (1988) The capacity of plastids from developing pea cotyledons to synthesise acetyl CoA. Planta 173, 172–182
Dry, I., Smith, A.M., Edwards, E.A., Bhattacharyya, M.K., Dunn, P., Martin, C. (1992) Characterisation of cDNAS encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs. Plant Journal, in press
Edwards, J., Green, J.G., ap Rees, T. (1988) Activity of branching enzyme as a cardinal feature of the Ra locus in Pisum sativum. Phytochemistry 27, 1615–1620
Goldner, W., Beevers, H. (1989) Starch synthase and starch branching enzyme from germinating castor bean endosperm. Phytochemistry 28, 1809–1812
Hawker, J.S., Ozbun, J.L., Preiss, J. (1972) Unprimed starch synthesis by soluble ADP glucose-starch glucosyltransferase from potato tubers. Phytochemistry 11, 1287–1293
Hawker, J.S., Ozbun, J.L., Ozaki, H., Greenberg, E., Preiss, J. (1974) Interaction of spinach leaf adenosine diphosphate glucose α-l,4-glucan α-4-glucosyl transferase and α-1,4-glucan, α-1,4-glucan-6-glycosyl transferase in synthesis of branched α-glucan. Arch. Biochem. Biophys. 160, 530–551
Krauss, A., Marschner, H. (1984) Growth rate and carbohydrate metabolism of potato tubers exposed to high temperatures. Potato Res. 27, 297–303
Kruger, N.J., ap Rees, T. (1983) Properties of α-glucan phosphorylase from pea chloroplasts. Phytochemistry 22, 1891–1898
Hedley, C.L., Smith, C.M., Ambrose, M.J., Cook, S., Wang, T.L. (1986) 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
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
MacDonald, F.D., Preiss, J. (1985) Partial purification and characterisation of granule-bound starch synthases from normal and Waxy maize. Plant Physiol. 78, 849–852
Matters, G.L., Boyer, C.D. (1981) Starch synthases and starch branching enzymes from Pisum sativum. Phytochemistry 20, 1804–1809
Matters, G.L., Boyer, C.D. (1982) Soluble starch synthases and starch branching enzymes from cotyledons of smooth-and wrinkled-seeded lines of Pisum sativum L. Biochem. Genet. 20, 833–848
Ozbun, J.L., Hawker, J.S., Preiss, J. (1971a) Adenosine diphosphoglucose-starch glucosyltransferases from developing kernels of waxy maize. Plant Physiol. 48, 765–769
Ozbun, J.L., Hawker, J.S., Preiss, J. (1971b) Multiple forms of α-1,4-glucan synthetase from spinach leaves. Biochem. Biophys. Res. Commun. 43, 631–636
Pollock, C., Preiss, J. (1980) The citrate-stimulated starch synthase of starchy maize kernels: purification and properties. Arch. Biochem. Biophys. 204, 578–588
Preiss, J. (1988) Biosynthesis of starch and its regulation. In: The biochemistry of plants, vol. 14, pp. 181–254, Preiss, J., ed. Academic Press, London
Rijven, A.H.G.C. (1986) Heat inactivation of starch synthase in wheat endosperm. Plant Physiol. 81, 448–453
Schiefer, S., Lee, E.Y.C., Whelan, W.J. (1978) The requirement for a primer in the in vitro synthesis of polysaccharide by sweetcorn (1–4)-α-d-glucan synthase. Carbohydr. Res. 61, 239–252
Schulman, A.H., Ahokas, H. (1990) A novel shrunken endosperm mutant of barley. Physiol. Plant. 78, 583–589
Shure, M., Wessier, S., Fedoroff, N. (1983) Molecular identification and isolation of the Waxy locus in maize. Cell 35, 225–233
Smith, A.M. (1988) Major differences in isoforms of starch-branching enzyme between developing embryos of round-and wrinkled-seeded peas (Pisum sativum L.). Planta 175, 270–279
Smith, A.M. (1990) Evidence that the ‘waxy’ protein of pea (Pisum sativum L.) starch granules is not the major, granule-bound starch synthase. Planta 182, 599–604
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We are grateful to Professor D.R. Davies, Professor T. ap Rees (Botany Department, University of Cambridge, UK), Dr. S. Raws-thorne, Dr. C. Hedley and Mr. L. Hill for helpful discussions during the preparation of this manuscript and to Dr. C. Hedley for the gift of the pea seeds. We would also like to thank the photography department and the glasshouse staff for their assistance and T. Dalzell for the typing.
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Denyer, K., Smith, A.M. The purification and characterisation of the two forms of soluble starch synthase from developing pea embryos. Planta 186, 609–617 (1992). https://doi.org/10.1007/BF00198043
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DOI: https://doi.org/10.1007/BF00198043