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. 1987 Apr;83(4):982–987. doi: 10.1104/pp.83.4.982

Partial Purification of Digitonin-Solubilized β-Glucan Synthase from Red Beet Root 1

Laura L Eiberger 1, Bruce P Wasserman 1
PMCID: PMC1056487  PMID: 16665376

Abstract

An enriched glucan synthase fraction was obtained from red beet root microsomes by sequential extraction with the detergents 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate and digitonin. The digitonin suspension was centrifuged on a glycerol gradient, where a glucan synthase peak with a specific activity of 30- to 40-fold over microsomes was recovered. Most protein contaminants were found in the gradient pellet. The glucan synthase-containing fraction was largely free of plasma membrane and tonoplast-derived ATPase activity and was enriched with a protein subunit of 68 kilodaltons.

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Selected References

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

  1. Addison R., Scarborough G. A. Solubilization and purification of the Neurospora plasma membrane H+-ATPase. J Biol Chem. 1981 Dec 25;256(24):13165–13171. [PubMed] [Google Scholar]
  2. Aloni Y., Cohen R., Benziman M., Delmer D. Solubilization of the UDP-glucose:1,4-beta-D-glucan 4-beta-D-glucosyltransferase (cellulose synthase) from Acetobacter xylinum. A comparison of regulatory properties with those of the membrane-bound form of the enzyme. J Biol Chem. 1983 Apr 10;258(7):4419–4423. [PubMed] [Google Scholar]
  3. Bennett A. B., O'neill S. D., Spanswick R. M. H-ATPase Activity from Storage Tissue of Beta vulgaris: I. Identification and Characterization of an Anion-Sensitive H-ATPase. Plant Physiol. 1984 Mar;74(3):538–544. doi: 10.1104/pp.74.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowman B. J., Blasco F., Slayman C. W. Purification and characterization of the plasma membrane ATPase of Neurospora crassa. J Biol Chem. 1981 Dec 10;256(23):12343–12349. [PubMed] [Google Scholar]
  5. Briskin D. P., Poole R. J. Characterization of a k-stimulated adenosine triphosphatase associated with the plasma membrane of red beet. Plant Physiol. 1983 Feb;71(2):350–355. doi: 10.1104/pp.71.2.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Delmer D. P., Heiniger U., Kulow C. UDP-glucose: Glucan Synthetase in Developing Cotton Fibers: I. Kinetic and Physiological Properties. Plant Physiol. 1977 Apr;59(4):713–718. doi: 10.1104/pp.59.4.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dufour J. P., Goffeau A. Solubilization by lysolecithin and purification of the plasma membrane ATPase of the yeast Schizosaccharomyces pombe. J Biol Chem. 1978 Oct 10;253(19):7026–7032. [PubMed] [Google Scholar]
  8. Henry R. J., Stone B. A. Solubilization of beta-glucan synthases from the membranes of cultured ryegrass endosperm cells. Biochem J. 1982 Jun 1;203(3):629–636. doi: 10.1042/bj2030629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hjelmeland L. M., Chrambach A. Solubilization of functional membrane proteins. Methods Enzymol. 1984;104:305–318. doi: 10.1016/s0076-6879(84)04097-0. [DOI] [PubMed] [Google Scholar]
  10. Kasamo K. Purification and Properties of the Plasma Membrane H-Translocating Adenosine Triphosphatase of Phaseolus mungo L. Roots. Plant Physiol. 1986 Apr;80(4):818–824. doi: 10.1104/pp.80.4.818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Mandala S., Taiz L. Partial purification of a tonoplast ATPase from corn coleoptiles. Plant Physiol. 1985 Jun;78(2):327–333. doi: 10.1104/pp.78.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mueller S. C., Brown R. M., Jr Evidence for an intramembrane component associated with a cellulose microfibril-synthesizing complex in higher plants. J Cell Biol. 1980 Feb;84(2):315–326. doi: 10.1083/jcb.84.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. O'neill S. D., Spanswick R. M. Effects of vanadate on the plasma membrane ATPase of red beet and corn. Plant Physiol. 1984 Jul;75(3):586–591. doi: 10.1104/pp.75.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Press G. A., Glazer H. S., Wasserman T. H., Aronberg D. J., Lee J. K., Sagel S. S. Thoracic wall involvement by Hodgkin disease and non-Hodgkin lymphoma: CT evaluation. Radiology. 1985 Oct;157(1):195–198. doi: 10.1148/radiology.157.1.4034966. [DOI] [PubMed] [Google Scholar]
  16. Péaud-Lenoël C., Axelos M. Structural features of the beta-glucans enzymatically synthesized from uridine diphosphate glucose by wheat seedlings. FEBS Lett. 1970 Jun 8;8(4):224–228. doi: 10.1016/0014-5793(70)80270-8. [DOI] [PubMed] [Google Scholar]
  17. Serrano R. Purification of the proton pumping ATPase from plant plasma membranes. Biochem Biophys Res Commun. 1984 Jun 15;121(2):735–740. doi: 10.1016/0006-291x(84)90243-2. [DOI] [PubMed] [Google Scholar]
  18. Smith M. M., Stone B. A. Beta-glucan synthesis by cell-free extracts from Lolium multiflorum endosperm. Biochim Biophys Acta. 1973 Jun 20;313(1):72–94. doi: 10.1016/0304-4165(73)90189-x. [DOI] [PubMed] [Google Scholar]
  19. Smith R., Scarborough G. A. Large-scale isolation of the Neurospora plasma membrane H+-ATPase. Anal Biochem. 1984 Apr;138(1):156–163. doi: 10.1016/0003-2697(84)90784-x. [DOI] [PubMed] [Google Scholar]
  20. Thelen M. P., Delmer D. P. Gel-Electrophoretic Separation, Detection, and Characterization of Plant and Bacterial UDP-Glucose Glucosyltransferases. Plant Physiol. 1986 Jul;81(3):913–918. doi: 10.1104/pp.81.3.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tsai C. M., Hassid W. Z. Solubilization and Separation of Uridine Diphospho-d-glucose: beta-(1 --> 4) Glucan and Uridine Diphospho-d-glucose:beta-(1 --> 3) Glucan Glucosyltransferases from Coleoptiles of Avena sativa. Plant Physiol. 1971 Jun;47(6):740–744. doi: 10.1104/pp.47.6.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Van Der Woude W. J., Lembi C. A., Morré D. J. beta-Glucan Synthetases of Plasma Membrane and Golgi Apparatus from Onion Stem. Plant Physiol. 1974 Sep;54(3):333–340. doi: 10.1104/pp.54.3.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wasserman B. P., McCarthy K. J. Regulation of Plasma Membrane beta-Glucan Synthase from Red Beet Root by Phospholipids : Reactivation of Triton X-100 Extracted Glucan Synthase by Phospholipids. Plant Physiol. 1986 Oct;82(2):396–400. doi: 10.1104/pp.82.2.396. [DOI] [PMC free article] [PubMed] [Google Scholar]

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