A novel biotinylated heterobifunctional cross-linking reagent bearing an aromatic diazirine.
The synthesis of a p-[(3-trifluoromethyl)diazirine-3-yl]benzoic acid derivative is described as a new carbene generating heterobifunctional cross-linking reagent. The cross-linker carries a biotin moiety in order to make use of avidin-biotin technology for specific manipulation of cross-linked components. To evaluate the ability of this reagent, the inter-subunit cross-linking of egg-white avidin tetramer was investigated. As a typical application of avidin-biotin technology for cross-linking experiments, a chemiluminescent detection method was examined to identify photobiotinylated components. A cross-linked dimeric product with an apparent molecular mass of 38 kDa was clearly visualized by the combined use of a horseradish peroxidase-streptavidin conjugate and a luminol-based chemiluminescent system.
Hatanaka
Y
Y
Research Institute for Wakan-Yaku, Toyama Medical and Pharmaceutical University, Japan.
Hashimoto
M
M
Kanaoka
Y
Y
D001360:Avidin / Q000737:chemistry; D001389:Azirines / Q000138:chemical synthesis* / Q000737:chemistry; D001710:Biotin / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008163:Luminescent Measurements; D015394:Molecular Structure; D010782:Photolysis; D013053:Spectrophotometry
D001389:Azirines; D003432:Cross-Linking Reagents; D001360:Avidin; D001710:Biotin