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A fluorescent radioiodinated oligonucleotidic photoaffinity probe for protein labeling: synthesis and photolabeling of thrombin.

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Berens, C Courtoy, Pierre J. [UCL] Sonveaux, Etienne [UCL]

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.

Document type Article de périodique (Journal article) – Journal Article, Research Support, Non-U.S. Gov't
Access type Accès restreint
Publication date 1999
Journal information "Bioconjugate chemistry" - Vol. 10, no. 1, p. 56-61 (1999)
Peer reviewed yes
issn 1043-1802
Publication status Publié
Affiliation UCL - MD/BICL - Département de biochimie et de biologie cellulaire
MESH Subject Animals ; Benzophenones - chemistry ; Cattle ; Electrophoresis, Polyacrylamide Gel ; Fluorescent Dyes - chemistry - radiation effects ; Humans ; Iodine Radioisotopes ; Microscopy, Confocal ; Oligodeoxyribonucleotides - chemistry - radiation effects ; Photoaffinity Labels - chemistry - radiation effects ; Proteins - chemistry ; Thrombin - chemistry - radiation effects ; Ultraviolet Rays
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Bibliographic reference Berens, C ; Courtoy, Pierre J. ; Sonveaux, Etienne. A fluorescent radioiodinated oligonucleotidic photoaffinity probe for protein labeling: synthesis and photolabeling of thrombin.. In: Bioconjugate chemistry, Vol. 10, no. 1, p. 56-61 (1999)
Permanent URL http://hdl.handle.net/2078.1/28773