Post-biotinylation of photocrosslinking by Staudinger-Bertozzi ligation of preinstalled alkylazide tag.
Post-biotinylation of the alkyl azide derivative of trifluoromethyl phenyldiazirine (TPD) was elucidated to apply a photoaffinity biotinylation technique. A photo-modified polyvinilidene difluoride (PVDF) membrane was used as a photolabeled component and we introduced biotin by Staudinger-Bertozzi ligation. The 15 pmol amount of biotinylated reagent was still effective for the visualization of cross-linked product on the matrix. The results show the potential utility of alkyl azide carrying TPD derivatives in the application of photoaffinity biotinylation, which could be useful for the ligands with tight structural requirements.
Hashimoto
Makoto
M
Department of Agricultural and Life Science, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan. hasimoto@obihiro.ac.jp
Hatanaka
Yasumaru
Y
D000478:Alkylation; D001386:Azides / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D003432:Cross-Linking Reagents; D006207:Half-Life; D009682:Magnetic Resonance Spectroscopy; D008567:Membranes, Artificial; D019920:Photoaffinity Labels; D010777:Photochemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013056:Spectrophotometry, Ultraviolet
D001386:Azides; D003432:Cross-Linking Reagents; D008567:Membranes, Artificial; D019920:Photoaffinity Labels; D001710:Biotin