The use of sulfosuccinimidyl-2-(p-azidosalicylamido)-1,3′-dithiopropionate as a crosslinking reagent to identify cell surface receptors

Enid G. Shephard, Frederick C. de Beer, Claus von Holt, Janet P. Hapgood

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Conditions for solubilizing and iodinating the heterobifunctional thiol-cleavable photoreactive crosslinking reagent sulfosuccinimidyl-2-(p-azidosalicylamido)-1,3′-dithiopropionate which leave the ester moiety, disulfide bond, and azido group reactive are described. Iodination was performed in a mixture of dimethyl sulfoxide and bicarbonate, pH 9.0 (1:20, v v), as solubilizing agent and Iodogen as oxidant. The lectin phytohemagglutinin was derivatized with the iodinated crosslinker and the interaction between phytohemagglutinin and mononuclear cells was chosen as the model system to monitor the efficiency of sulfosuccinimidyl-2-(p-azidosalicylamido)-1,3′-dithiopropionate as a crosslinking reagent. Transfer of 125I to the biologically significant T11 lymphocyte receptor in addition to 125I labeling of other membrane proteins to which the lectin binds was detected by polyacrylamide gel electrophoresis under reducing conditions.

Original languageEnglish
Pages (from-to)306-313
Number of pages8
JournalAnalytical Biochemistry
Volume168
Issue number2
DOIs
StatePublished - Feb 1 1988

Bibliographical note

Funding Information:
This work was supported by grants from the South African Medical Research Council (Frederick C. de Beer) and the Foundation for Research Development (Claus von Holt). S. Beer is thanked for technical assistance.

Keywords

  • cell membrane receptors
  • cleavable crosslinking
  • iodination
  • photoaffinity label
  • phytohemagglutinin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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