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. 1993 Dec;103(4):1355–1359. doi: 10.1104/pp.103.4.1355

Biochemical Evidence for the Role of the Waxy Protein from Pea (Pisum sativum L.) as a Granule-Bound Starch Synthase.

M N Sivak 1, M Wagner 1, J Preiss 1
PMCID: PMC159126  PMID: 12232029

Abstract

Proteins were solubilized from starch extracted from developing pea (Pisum sativum L.) embryos and chromatography of these proteins on a Mono-Q column separated two peaks of starch synthase activity. The major activity peak comprised more than 80% of the total activity. This fraction contained only the Waxy protein, as shown by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate followed by staining for proteins or by immunoblot. A 77-kD polypeptide associated with the starch granules and presumed by others to be a starch synthase could not be detected in any of the active fractions. The native molecular weight of the solubilized starch synthase was 59,600 [plus or minus] 1700 as determined by sucrose density gradient. It is concluded that in pea seeds the Waxy protein and the starch synthase bound to the granule are the same protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  2. Dry I., Smith A., Edwards A., Bhattacharyya M., Dunn P., Martin C. Characterization of cDNAs encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs of pea and potato. Plant J. 1992 Mar;2(2):193–202. [PubMed] [Google Scholar]
  3. Ghosh H. P., Preiss J. Adenosine diphosphate glucose pyrophosphorylase. A regulatory enzyme in the biosynthesis of starch in spinach leaf chloroplasts. J Biol Chem. 1966 Oct 10;241(19):4491–4504. [PubMed] [Google Scholar]
  4. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  5. Lovell S. J., Winzor D. J. Effects of phosphate on the dissociation and enzymic stability of rabbit muscle lactate dehydrogenase. Biochemistry. 1974 Aug 13;13(17):3527–3531. doi: 10.1021/bi00714a018. [DOI] [PubMed] [Google Scholar]
  6. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  7. Macdonald F. D., Preiss J. Partial purification and characterization of granule-bound starch synthases from normal and waxy maize. Plant Physiol. 1985 Aug;78(4):849–852. doi: 10.1104/pp.78.4.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. NELSON O. E., RINES H. W. The enzymatic deficiency in the waxy mutant of maize. Biochem Biophys Res Commun. 1962 Oct 31;9:297–300. doi: 10.1016/0006-291x(62)90043-8. [DOI] [PubMed] [Google Scholar]
  9. Nelson O. E., Chourey P. S., Chang M. T. Nucleoside Diphosphate Sugar-Starch Glucosyl Transferase Activity of wx Starch Granules. Plant Physiol. 1978 Sep;62(3):383–386. doi: 10.1104/pp.62.3.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Noyes B. E., Glatthaar B. E., Garavelli J. S., Bradshaw R. A. Structural and functional similarities between mitochondrial malate dehydrogenase and L-3-hydroxyacyl CoA dehydrogenase. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1334–1338. doi: 10.1073/pnas.71.4.1334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  12. Visser R. G., Somhorst I., Kuipers G. J., Ruys N. J., Feenstra W. J., Jacobsen E. Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs. Mol Gen Genet. 1991 Feb;225(2):289–296. doi: 10.1007/BF00269861. [DOI] [PubMed] [Google Scholar]

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