Premium
An Isotope‐Coded Fluorogenic Cross‐Linker for High‐Performance Target Identification Based on Photoaffinity Labeling
Author(s) -
Tomohiro Takenori,
Morimoto Shota,
Shima Toshiya,
Chiba Junya,
Hatanaka Yasumaru
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201408580
Subject(s) - moiety , photoaffinity labeling , chemistry , linker , mass spectrometry , fluorescence , combinatorial chemistry , identification (biology) , computational biology , biochemistry , chromatography , binding site , stereochemistry , computer science , biology , physics , botany , quantum mechanics , operating system
A photoaffinity labeling (PAL)‐based method for the rapid identification of target proteins is presented in which a high‐performance chemical tag, an isotope‐coded fluorescent tag (IsoFT), can be attached to the interacting site by irradiation. Labeled peptides can be easily distinguished among numerous proteolytic digests by sequential detection with highly sensitive fluorescence spectroscopy and mass spectrometry. Subsequent MS/MS analysis provides amino acid sequence information with a higher depth of coverage. The combination of PAL and heterogeneous target‐selecting techniques significantly reduces the amount of time and protein required for identification. An additional photocleavable moiety successfully accelerated proteomic analysis using cell lysate. This method is a widely applicable approach for the rapid and accurate identification of interacting proteins.