Application of 3-[3-(3-(trifluoromethyl)diazirin-3-yl)phenyl]-2,3-dihydroxypropionic acid, carbene-generating, cleavable cross-linking reagent for photoaffinity labeling

✍ Scribed by Dmitry E. Bochkariov; Alexander A. Kogon

• Book ID: 102988768
• Publisher: Elsevier Science
• Year: 1992
• Tongue: English
• Weight: 716 KB
• Volume: 204
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(92)90144-v

✦ Synopsis

N-Hydroxysuccinimide ester of 3-[3-(3-(trifluoromethyl)diazirin-3-yl)phenyl]-2,3-dihydroxypro pionic acid was successfully tested in a ribosomal tRNA binding system. It is an originally designed trifluoromethyl-diazirine-based cleavable cross-linking reagent with a very short distance between the active points (about 8.5 A). The reagent was coupled to the amino acid amino group of Phe-tRNAPhe to obtain a photoactivatable analog of peptidyl-tRNA. This analog was bound to ribosomes and the complex was irradiated with uv light. After isolation, the cross-linked product was cleft by periodate treatment to reveal the properties of the new reagent.