Application of 3-[3-(3-(trifluoromethyl)diazirin-3-yl)phenyl]-2,3- dihydroxypropionic acid, carbene-generating, cleavable cross-linking reagent for photoaffinity labeling.
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.
Bochkariov
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DE
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow region.
Kogon
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AA
D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry; D001388:Aziridines / Q000138:chemical synthesis / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D006838:Hydrocarbons; D008697:Methane / Q000031:analogs & derivatives / Q000138:chemical synthesis; D015394:Molecular Structure; D012329:RNA, Bacterial / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000378:metabolism; D012360:RNA, Transfer, Phe; D012270:Ribosomes / Q000378:metabolism; D013388:Succinimides / Q000138:chemical synthesis / Q000737:chemistry; D014466:Ultraviolet Rays
D000345:Affinity Labels; D001388:Aziridines; D003432:Cross-Linking Reagents; D006838:Hydrocarbons; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D012360:RNA, Transfer, Phe; D013388:Succinimides; C076403:3-(3-(3-(trifluoromethyl)diazirin-3-yl)phenyl)-2,3-dihydroxypropionic acid N-hydroxysuccinimide ester; C030011:carbene; D008697:Methane