Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Jul;159(1):53–56. doi: 10.1128/jb.159.1.53-56.1984

Isolation and partial characterization of a carbohydrate-binding protein from a nematode-trapping fungus.

C A Borrebaeck, B Mattiasson, B Nordbring-Hertz
PMCID: PMC215591  PMID: 6539773

Abstract

A developmentally regulated carbohydrate-binding protein from the capture organs of Arthrobotrys oligospora, not present on hyphae, was isolated and partially characterized. Surface structures of A. oligospora were radiolabeled with [125I]iodosulfanilic acid. The fungus was homogenized, and the homogenate was passed over an affinity column containing N-acetyl-D-galactosamine immobilized to Sepharose 6B. The bound radiolabeled protein was eluted from the affinity column with a glycine-hydrochloride buffer (pH 3.0), concentrated, and chromatographed on a metal chelate affinity gel containing Ca2+. EDTA was used as an eluant for the radiolabeled protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with autoradiography revealed a molecular weight for the carbohydrate- and cation-binding polypeptide of ca. 20,000.

Full text

PDF
53

Images in this article

Selected References

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

  1. Borrebaeck C. A., Lönnerdal B., Etzler M. E. Metal ion content of dolichos biflorus lectin and effect of divalent cations on lectin activity. Biochemistry. 1981 Jul 7;20(14):4119–4122. doi: 10.1021/bi00517a026. [DOI] [PubMed] [Google Scholar]
  2. Carraway K. L. Covalent labeling of membranes. Biochim Biophys Acta. 1975 Dec 29;415(4):379–410. doi: 10.1016/0304-4157(75)90005-2. [DOI] [PubMed] [Google Scholar]
  3. Eschenbruch M., Bürk R. R. Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal Biochem. 1982 Sep 1;125(1):96–99. doi: 10.1016/0003-2697(82)90387-6. [DOI] [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. Porath J., Carlsson J., Olsson I., Belfrage G. Metal chelate affinity chromatography, a new approach to protein fractionation. Nature. 1975 Dec 18;258(5536):598–599. doi: 10.1038/258598a0. [DOI] [PubMed] [Google Scholar]
  6. Sears D. A., Reed C. F., Helmkamp R. W. A radioactive label for the erythrocyte membrane. Biochim Biophys Acta. 1971 Jun 1;233(3):716–719. doi: 10.1016/0005-2736(71)90170-2. [DOI] [PubMed] [Google Scholar]
  7. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES