Selective detection of 2-nitrobenzenesulfenyl-labeled peptides by matrix-assisted laser desorption/ionization-time of flight mass spectrometry using a novel matrix

Proteomics
Ei-Ichi MatsuoOsamu Nishimura

Abstract

The 2-nitrobenzenesulfenyl (NBS) method, which is useful for quantitative proteome analysis, is based on stable isotope labeling of tryptophan residues with NBS chloride ((12)C(6)-NBSCl or (13)C(6)-NBSCl). We found that 3-hydroxy-4-nitrobenzoic acid (3H4NBA) is a more suitable matrix than 2,5-dihydroxybenzoic acid (DHB) for detecting NBS-labeled peptides by MALDI-quadrupole IT (QIT)-TOF MS . Furthermore, NBS-labeled peptides were selectively ionized and detected in a mixture of NBS-labeled and unlabeled peptides. Labeled paired peaks were easily detected without enrichment, nonpaired labeled peaks were clearly distinguished from unlabeled contaminating peptides, and nitrotyrosine-containing peptides were also selectively detected on the 3H4NBA matrix, while by-product-peaks arising from nitrobenzene moieties were suppressed. The use of 3H4NBA as a comatrix with CHCA improved the sensitivity of detection while substantially retaining the selectivity of 3H4NBA. The 3H4NBA matrix offers great advantages in terms of simplicity, sensitivity, and usability when used for the NBS method and for MALDI-TOF MS analysis applied to compounds having a nitrobenzene ring.

References

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Nov 26, 2005·Rapid Communications in Mass Spectrometry : RCM·Ei-Ichi MatsuoOsamu Nishimura

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Citations

Nov 7, 2006·Analytical and Bioanalytical Chemistry·Chensong PanHanfa Zou
Jun 19, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ei-ichi MatsuoOsamu Nishimura
Dec 13, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Catherine Fenselau
Jan 3, 2007·Journal of Mass Spectrometry : JMS
Jun 13, 2012·Journal of Mass Spectrometry : JMS·Toni TodorovskiRalf Hoffmann
Mar 20, 2010·Hepatology Research : the Official Journal of the Japan Society of Hepatology·Yoichiro TakamiHirohito Tsubouchi
Jun 16, 2016·Plant Signaling & Behavior·Misa TakahashiHiromichi Morikawa
Jul 29, 2016·Analytical Chemistry·Yuko FukuyamaKoichi Tanaka

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