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Charge derivatization by 4-sulfophenyl isothiocyanate enhances peptide sequencing by post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

✍ Scribed by Lyuben N. Marekov; Peter M. Steinert

Publisher
John Wiley and Sons
Year
2003
Tongue
English
Weight
106 KB
Volume
38
Category
Article
ISSN
1076-5174

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✦ Synopsis

Abstract

High‐sensitivity, rapid identification of proteins in proteomic studies normally uses a combination of one‐ or two‐dimensional electrophoresis together with mass spectrometry. The simplicity and sensitivity of matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) have increased its application in recent years. The most common method of ‘peptide fingerprinting’ often may not provide robust identification. Normally additional sequence information by post‐source decay (PSD) MALDI‐TOFMS provides additional constraints for database searches to achieve highly confident results. Here we describe a derivatization procedure to facilitate the acquisition of such sequence information. Peptide digests from a skin‐expressed protein were modified with 4‐sulfophenyl isothiocyanate. The resulting peptides carry a fixed negative charge at the N‐terminal end and the resulting PSD spectrum is dominated by C‐terminal y‐type ions. The sequence information in most cases can be obtained manually or with simple programming tools. Methods of optimizing the procedure and increasing the sensitivity are discussed. Copyright © 2003 John Wiley & Sons, Ltd.

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