Binding position analysis of target proteins with the use of amidopyrene probes as LA-LDI enhancing tags. Issue 36 (22nd August 2016)

• Record Type: Journal Article
• Authors: Yoneda, Kozo; Hu, Yaping; Watanabe, Rei; Kita, Masaki; Kigoshi, Hideo
• Is Part Of: Organic & biomolecular chemistry. Volume 14:Issue 36(2016)
• Page Start: 8564
• Page End: 8569
• Publication Date: 2016-08-22
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/c6ob01381c
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : Selective detection of amidopyrene-labeled peptides by LA-LDI MS enabled us to analyze the binding position of ligands on target proteins. Abstract : Amidopyrene-conjugated compounds can be detected by label-assisted laser desorption/ionization mass spectrometry (LA-LDI MS) without matrixes. When actin, a cytoskeletal protein, was labeled with an excess amount of amidopyrene N -hydroxysuccinate (apy–OSu), eight apy-labeled actin peptides were predominantly detected by LA-LDI MS. Then actin was labeled with an amidopyrene NHS ester of the antitumor marine macrolide aplyronine A (ApA–apy–OSu) to form a 1 : 1 conjugate. The sequence of an apy-labeled peptide was established as A 108 PLNPKANR 116 by MS/MS analysis, in which the NHS ester moiety specifically reacted with the ε-amino group of K113. While the fragmentation at the linker part reduces the detection sensitivity of apy-labeled peptides on LA-LDI MS, our chemical probe method is useful for analyzing the binding modes of various ligands and target biomacromolecules that include multiple and weak interactions.