3-Diazirine-derivatives of bile salts for photoaffinity labeling.
New carbene-generating photolabile bile salt derivatives, 3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta [7 beta-3H]cholan-24-oic acid and (3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta [7 beta-3H]cholan-24-oyl)-2- aminoethanesulfonic acid were synthesized with high specific radioactivity. These 3-diazirine-derivatives could be activated to the corresponding carbenes by irradiation with ultraviolet light at 350 nm with a half-life time of 2 min. The 3-diazirine derivatives behaved in enterohepatic circulation like the natural bile salts. The uptake of [3H]taurocholate into isolated hepatocytes was competitively inhibited by (3,3-azo-7 alpha,12 alpha-dihydroxy-5 beta-cholan-24-oyl)-2- aminoethanesulfonic acid indicating that the 3,3-azo-derivative of taurocholate shares the hepatic transport systems for natural bile salts. It was demonstrated that the radioactively labeled 3-diazirine bile salt derivatives are useful probes for photoaffinity labeling of bile salt binding proteins especially in intact cells and tissues.
Kramer
W
W
Institut für Organische Chemie und Biochemie, Universität Freiburg, Federal Republic of Germany.
Schneider
S
S
D000345:Affinity Labels / Q000138:chemical synthesis*; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism*; D001692:Biological Transport; D003978:Diazomethane; D003979:Diazonium Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D004764:Enterohepatic Circulation; D008099:Liver / Q000378:metabolism; D010777:Photochemistry; D051381:Rats
D000345:Affinity Labels; D001647:Bile Acids and Salts; D003979:Diazonium Compounds; D003978:Diazomethane