Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1992 Sep;100(1):457–463. doi: 10.1104/pp.100.1.457

Partial Purification and Properties of an Inducible Uridine 5′-Diphosphate-Glucose-Salicylic Acid Glucosyltransferase from Oat Roots 1

Nasser Yalpani 1,2, Margot Schulz 1,3, Michael P Davis 1,4, Nelson E Balke 1
PMCID: PMC1075572  PMID: 16652983

Abstract

A salicylic acid (SA)-inducible uridine 5′-diphosphate (UDP)-glucose:SA 3-O-glucosyltransferase was extracted from oat (Avena sativa L. cv Dal) roots. Reverse phase high-performance liquid chromatography or anion exchange chromatography was used to separate SA from the product, β-O-d-glucosylsalicylic acid. The soluble enzyme was purified 176-fold with 5% recovery using a combination of pH fractionation, anion exchange, gel filtration, and chromatofocusing chromatography. The partially purified protein had a native molecular weight of about 50,000, an apparent isoelectric point at pH 5.0, and maximum activity at pH 5.5. The enzyme had a Km of 0.28 mm for UDP-glucose and was highly specific for this sugar donor. More than 20 hydroxybenzoic and hydroxycinnamic acid derivatives were assayed as potential glucose acceptors. UDP-glucose:SA 3-O-glucosyltransferase activity was highly specific toward SA (Km = 0.16 mm). The enzyme was inhibited by UDP and uridine 5′-triphosphate but not by up to 7.5 mm uridine 5′-monophosphate.

Full text

PDF
462

Images in this article

Selected References

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

  1. Bajaj K. L., de Luca V., Khouri H., Ibrahim R. K. Purification and Properties of Flavonol-Ring B Glucosyltransferase from Chrysosplenium americanum. Plant Physiol. 1983 Jul;72(3):891–896. doi: 10.1104/pp.72.3.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bechthold A., Berger U., Heide L. Partial purification, properties, and kinetic studies of UDP-glucose:p-hydroxybenzoate glucosyltransferase from cell cultures of Lithospermum erythrorhizon. Arch Biochem Biophys. 1991 Jul;288(1):39–47. doi: 10.1016/0003-9861(91)90162-c. [DOI] [PubMed] [Google Scholar]
  3. Enyedi A. J., Yalpani N., Silverman P., Raskin I. Localization, conjugation, and function of salicylic acid in tobacco during the hypersensitive reaction to tobacco mosaic virus. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2480–2484. doi: 10.1073/pnas.89.6.2480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Glass A. D. Influence of phenolic acids on ion uptake: I. Inhibition of phosphate uptake. Plant Physiol. 1973 Jun;51(6):1037–1041. doi: 10.1104/pp.51.6.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Harper J. R., Balke N. E. Characterization of the inhibition of k absorption in oat roots by salicylic Acid. Plant Physiol. 1981 Dec;68(6):1349–1353. doi: 10.1104/pp.68.6.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hurkman W. J., Tanaka C. K. Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol. 1986 Jul;81(3):802–806. doi: 10.1104/pp.81.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Leslie C. A., Romani R. J. Inhibition of ethylene biosynthesis by salicylic Acid. Plant Physiol. 1988 Nov;88(3):833–837. doi: 10.1104/pp.88.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Malamy J., Carr J. P., Klessig D. F., Raskin I. Salicylic Acid: a likely endogenous signal in the resistance response of tobacco to viral infection. Science. 1990 Nov 16;250(4983):1002–1004. doi: 10.1126/science.250.4983.1002. [DOI] [PubMed] [Google Scholar]
  9. Métraux J. P., Signer H., Ryals J., Ward E., Wyss-Benz M., Gaudin J., Raschdorf K., Schmid E., Blum W., Inverardi B. Increase in salicylic Acid at the onset of systemic acquired resistance in cucumber. Science. 1990 Nov 16;250(4983):1004–1006. doi: 10.1126/science.250.4983.1004. [DOI] [PubMed] [Google Scholar]
  10. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  11. Raskin I., Ehmann A., Melander W. R., Meeuse B. J. Salicylic Acid: a natural inducer of heat production in arum lilies. Science. 1987 Sep 25;237(4822):1601–1602. doi: 10.1126/science.237.4822.1601. [DOI] [PubMed] [Google Scholar]
  12. Shimizu T., Kojima M. Partial purification and characterization of UDPG:t-cinnamate glucosyltransferase in the root of sweet potato, Ipomoea batatas Lam. J Biochem. 1984 Jan;95(1):205–212. doi: 10.1093/oxfordjournals.jbchem.a134586. [DOI] [PubMed] [Google Scholar]
  13. Stitt M., Wirtz W., Heldt H. W. Metabolite levels during induction in the chloroplast and extrachloroplast compartments of spinach protoplasts. Biochim Biophys Acta. 1980 Nov 5;593(1):85–102. doi: 10.1016/0005-2728(80)90010-9. [DOI] [PubMed] [Google Scholar]
  14. Sun Y., Hrazdina G. Isolation and Characterization of a UDPGlucose: Flavonol O-Glucosyltransferase from Illuminated Red Cabbage (Brassica oleracea cv Red Danish) Seedlings. Plant Physiol. 1991 Feb;95(2):570–576. doi: 10.1104/pp.95.2.570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Yalpani N., Silverman P., Wilson T. M., Kleier D. A., Raskin I. Salicylic acid is a systemic signal and an inducer of pathogenesis-related proteins in virus-infected tobacco. Plant Cell. 1991 Aug;3(8):809–818. doi: 10.1105/tpc.3.8.809. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES