Efficient approach for profiling photoaffinity labeled peptides with a cleavable biotinyl photoprobe.
Based on the application of our recent biotinyl photoprobe with a cleavable N-acylsulfonamide, an efficient process has been developed for profiling photoaffinity labeled peptides among a large excess of unlabeled concomitants. N-acylsulfonamide group was found to be stable under the usual S-pyridylethylation condition of cysteine residues whereas the group was easily cleaved by N-alkylation with iodoacetic acid in acidic condition. The selective nature between two common protein alkylation reactions was evaluated with L-glutamate dehydrogenase (GDH) using an acidic amino acid photoprobe with biotinylated acylsulfonamide function. The labeled GDH was successfully subjected to S-pyridylethylation keeping the biotin tag intact, and then was easily released from streptavidin matrix with high purity via iodoacetic acid-mediated alkylation under mild condition at pH 5.0.
Bongo
Nlandu B
NB
Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
Tomohiro
Takenori
T
Hatanaka
Yasumaru
Y
D000478:Alkylation; D001710:Biotin / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006868:Hydrolysis; D010455:Peptides / Q000737:chemistry*; D019920:Photoaffinity Labels; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010455:Peptides; D019920:Photoaffinity Labels; D001710:Biotin; D005969:Glutamate Dehydrogenase