N,N'-Bis(4-azidobenzoyl)cystine--a cleavable photoaffinity reagent

Anal Biochem. 1985 Sep;149(2):331-8. doi: 10.1016/0003-2697(85)90578-0.

Abstract

A water-soluble, cleavable, heterobifunctional photoaffinity label has been synthesized in one step from N-hydroxysuccinimidyl 4-azidobenzoate and cystine. The resultant compound, N,N'-bis(4-azidobenzoyl) cystine [(ABC)2], reacts with protein sulfhydryl groups through disulfide exchange to generate photoactive derivatives. Since [35S]cystine of high specific activity is readily available, it is possible to produce highly radioactive (ABC)2. ABC-derivatized ovalbumin is antigenic in vivo, and monoclonal antibodies specific for ABC have been produced. The antigen binding site of these antibodies was covalently labeled with ABC.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Affinity Labels / chemical synthesis*
  • Affinity Labels / immunology
  • Antibodies, Monoclonal / analysis
  • Antibody Specificity
  • Azides / chemical synthesis*
  • Azides / immunology
  • Cystine / analogs & derivatives*
  • Cystine / chemical synthesis
  • Cystine / immunology
  • Disulfides
  • Haptens
  • Photochemistry
  • Proteins / analysis
  • Solubility
  • Spectrophotometry, Ultraviolet
  • Sulfhydryl Compounds

Substances

  • Affinity Labels
  • Antibodies, Monoclonal
  • Azides
  • Disulfides
  • Haptens
  • Proteins
  • Sulfhydryl Compounds
  • Cystine
  • N,N'-bis(4-azidobenzoyl)cystine