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. 1992 Sep;100(1):397–402. doi: 10.1104/pp.100.1.397

Purification of an Infection-Related, Extracellular Peroxidase from Barley 1

Kent Kerby 1,2,2, Shauna C Somerville 1,2
PMCID: PMC1075564  PMID: 16652974

Abstract

Increases in two extracellular peroxidases were observed following inoculation of barley (Hordeum vulgare L.) with the powdery mildew pathogen (Erysiphe graminis DC.: Fr. f. sp. hordei Em. Marchal). The more prominent isozyme, P8.5, was purified from intercellular wash fluids by acetone precipitation, ion-exchange chromatography, isoelectric focusing, and gel filtration. Purified P8.5 is a heme-containing, glycoprotein with a Mr of 35,000. It has eight cysteine residues. A highly specific, high-titer antiserum to deglycosylated P8.5 was produced.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Hu C. F., van Huystee R. B. Role of carbohydrate moieties in peanut (Arachis hypogaea) peroxidases. Biochem J. 1989 Oct 1;263(1):129–135. doi: 10.1042/bj2630129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Kieliszewski M. J., Leykam J. F., Lamport D. T. Structure of the Threonine-Rich Extensin from Zea mays. Plant Physiol. 1990 Feb;92(2):316–326. doi: 10.1104/pp.92.2.316. [DOI] [PMC free article] [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. Li Z. C., McClure J. W., Hagerman A. E. Soluble and Bound Apoplastic Activity for Peroxidase, beta-d-Glucosidase, Malate Dehydrogenase, and Nonspecific Arylesterase, in Barley (Hordeum vulgare L.) and Oat (Avena sativa L.) Primary Leaves. Plant Physiol. 1989 May;90(1):185–190. doi: 10.1104/pp.90.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Mazza G., Welinder K. G. Covalent structure of turnip peroxidase 7. Cyanogen bromide fragments, complete structure and comparison to horseradish peroxidase C. Eur J Biochem. 1980 Jul;108(2):481–489. doi: 10.1111/j.1432-1033.1980.tb04745.x. [DOI] [PubMed] [Google Scholar]
  7. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  8. Rasmussen S. K., Welinder K. G., Hejgaard J. cDNA cloning, characterization and expression of an endosperm-specific barley peroxidase. Plant Mol Biol. 1991 Feb;16(2):317–327. doi: 10.1007/BF00020562. [DOI] [PubMed] [Google Scholar]
  9. Rebmann G., Hertig C., Bull J., Mauch F., Dudler R. Cloning and sequencing of cDNAs encoding a pathogen-induced putative peroxidase of wheat (Triticum aestivum L.). Plant Mol Biol. 1991 Feb;16(2):329–331. doi: 10.1007/BF00020563. [DOI] [PubMed] [Google Scholar]
  10. Saeki K., Ishikawa O., Fukuoka T., Nakagawa H., Kai Y., Kakuno T., Yamashita J., Kasai N., Horio T. Barley leaf peroxidase: purification and characterization. J Biochem. 1986 Feb;99(2):485–494. doi: 10.1093/oxfordjournals.jbchem.a135503. [DOI] [PubMed] [Google Scholar]
  11. Shannon L. M., Kay E., Lew J. Y. Peroxidase isozymes from horseradish roots. I. Isolation and physical properties. J Biol Chem. 1966 May 10;241(9):2166–2172. [PubMed] [Google Scholar]

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