Heterobifunctional Cross-Linking Agents Incorporating Perfluorinated Aryl Azides

Peter J. Crocker, Nobuyuki Imai, Krishnan Rajagopalan, Michael A. Boggess, Stefan Kwiatkowski, Lori D. Dwyer, Thomas C. Vanaman, David S. Watt

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

New heterobifunctional cross-linking reagents were developed that possess a photoactive tetrafluorinated phenyl azide as the photoactive terminus and a chemically reactive succinimidyl ester as the electrophilic terminus. These reagents, succinimidyl N-(4-azido-2,3,5,6-tetrafluorobenzoyl)tyrosinate (9) and succinimidyl 2-(4-azido-2,3,5,6-tetrafluorophenyl)thiazole-4-carboxylate (15), were designed to possess either an 125I or 35S radiolabel, respectively. In a biochemical study, the latter reagent was coupled to Lys-75 of calmodulin (CaM), and the radioiodinated monoadduct was photochemically cross-linked, in a calcium-dependent manner, to the porcine erythrocyte plasma membrane Ca2+,Mg2+-ATPase. Densitometry scans of the gel indicated a reproducible 22 % cross-linking of the CaM with one of the Ca2+,-Mg2+-ATPase bands. Since the purification of the Ca2+,Mg2+-ATPase results in micelles having Ca2+,-Mg2+-ATPase with its CaM binding site oriented both to the inside and outside of the micelle, the amount of Ca2+,Mg2+-ATPase available for cross-linking was reduced by approximately half, suggesting that the actual cross-linking efficiency was on the order of 40%.

Original languageEnglish
Pages (from-to)419-424
Number of pages6
JournalBioconjugate Chemistry
Volume1
Issue number6
DOIs
StatePublished - Nov 1 1990

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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