A fluorescent radioiodinated oligonucleotidic photoaffinity probe for protein labeling: synthesis and photolabeling of thrombin.
To study the interactions between oligonucleotides and proteins, an original photoaffinity radiolabeling probe has been synthesized. Starting with a 5'-pyridyldithio-3'-amino-oligonucleotide, the photophore benzophenone was first coupled to the 3' end, through acylation by an activated ester of benzoylbenzoic acid. A fluorescein molecule was grafted by alkylation of the free 5'-SH. This compound was finally radiolabeled with 125I using IodoBeads. The selective photolabeling of thrombin in a complex protein mixture by the radioiodinated probe validates this strategy to identify oligonucleotide-binding proteins.
Berens
C
C
Laboratoire de Chimie Thérapeutique et de Radiopharmacie, UCL 7340, 73 Avenue Emmanuel, Mounier, B-1200 Bruxelles, Belgium.
Courtoy
P J
PJ
Sonveaux
E
E
D000818:Animals; D001577:Benzophenones / Q000737:chemistry; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000528:radiation effects; D006801:Humans; D007457:Iodine Radioisotopes; D018613:Microscopy, Confocal; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000528:radiation effects; D019920:Photoaffinity Labels / Q000737:chemistry* / Q000528:radiation effects; D011506:Proteins / Q000737:chemistry*; D013917:Thrombin / Q000737:chemistry* / Q000528:radiation effects; D014466:Ultraviolet Rays
D001577:Benzophenones; D005456:Fluorescent Dyes; D007457:Iodine Radioisotopes; D009838:Oligodeoxyribonucleotides; D019920:Photoaffinity Labels; D011506:Proteins; C047723:benzophenone; D013917:Thrombin