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Efficient identification and quantification of proteins using isotope‐coded 1‐(6‐methylnicotinoyloxy)succinimides by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Tsumoto Hiroki,
Murata Chie,
Miyata Naoki,
Kohda Kohfuku,
Taguchi Ryo
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3279
Subject(s) - chemistry , reagent , derivatization , mass spectrometry , chromatography , peptide , matrix assisted laser desorption/ionization , succinimides , desorption , organic chemistry , biochemistry , adsorption
We describe a convenient and useful method for the identification and relative quantification of proteins using light and heavy reagents, 1‐(6‐methylnicotinoyloxy)succinimides (6‐CH 3 ‐Nic‐NHS and 6‐CD 3 ‐Nic‐NHS, respectively). This method is based on the chemical derivatization of amino groups of tryptic peptides with these reagents, i.e., the basic moiety of the reagents thus incorporated into both the N‐terminal amino group and the ε ‐amino group of the lysine residue would improve the ionization efficiency of tryptic peptides. An increase in protein sequence coverage is achieved by derivatization with these reagents or by combination of mass values before and after derivatization. Since a combination of 6‐CH 3 ‐Nic‐NHS and d 3 ‐labeled reagent (6‐CD 3 ‐Nic‐NHS) generates a 3 Da mass difference per reaction site, the d 3 ‐labeled reagent shifts the mass values of d 0 ‐labeled peptides according to the number of reactive amino groups in the peptides. In the case of tryptic peptides, the mass values of C‐terminal arginine and lysine peptides are shifted by 3 and 6 Da, respectively. Further, the 3 Da mass difference between 6‐CH 3 ‐Nic‐NHS and 6‐CD 3 ‐Nic‐NHS offers a means for the relative quantification of protein by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry. Copyright © 2007 John Wiley & Sons, Ltd.