Abstract
In studies of photoaffinity labeling, combinatorial use of mass spectrometry (MS) analysis has been increasing. Among the increasing numbers of such reports, however, major use of the method is for mapping labeled peptides, and application for identifying photolabeled peptides and their labeled amino acid residues is still limited. This is mainly caused that separation of the labeled peptide fragment(s) by HPLC, for example, is tedious and labour intensive. Therefore, in order to locate the ligand binding sites precisely even when photolabeling efficiency is low, a general and an efficient method to separate the photolabeled peptide(s) is required for identifying not only photolabeled peptides but also their labeled amino acid residues. As an example to fulfil this requirement, we here describe a method using simple purification step using anti-ligand antibody followed by MS analysis to characterize efficiently the binding sites for a calcium antagonist semotiadil in human serum albumin as the drug-binding protein.
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Kawahara, K., Masuda, K., Ishiguro, M., Nakayama, H. (2017). Utilities of Anti-ligand Antibody and Mass Spectrum to Elucidate Photolabeled Sites. In: Hatanaka, Y., Hashimoto, M. (eds) Photoaffinity Labeling for Structural Probing Within Protein. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56569-7_4
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DOI: https://doi.org/10.1007/978-4-431-56569-7_4
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