Protein identification by peptide mass fingerprinting and peptide sequence tagging with alternating scans of nano-liquid chromatography/infrared multiphoton dissociation Fourier transform ion cyclotron resonance mass spectrometry

✍ Scribed by Toshiyuki Kosaka; Tomoko Yoneyama-Takazawa; Kazuishi Kubota; Tatsuji Matsuoka; Ikuya Sato; Takamitsu Sasaki; Yorihisa Tanaka

• Publisher: John Wiley and Sons
• Year: 2003
• Tongue: English
• Weight: 334 KB
• Volume: 38
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/jms.545

✦ Synopsis

Abstract

We have developed a method for protein identification with peptide mass fingerprinting and sequence tagging using nano liquid chromatography (LC)/Fourier transform ion cyclotron resonance mass spectrometry (FTICR‐MS). To achieve greater sensitivity, a nanoelectrospray (nano‐ES) needle packed with reversed‐phase medium was used and connected to the nano‐ES ion source of the FTICR mass spectrometer. To obtain peptide sequence tag information, infrared multiphoton dissociation (IRMPD) was carried out in nano‐LC/FTICR‐MS analysis. The analysis involves alternating nano‐ES/FTICR‐MS and nano‐ES/IRMPD‐FTICR‐MS scans during a single LC run, which provides sets of parent and fragment ion masses of the proteolytic digest. The utility of this alternating‐scan nano‐LC/IRMPD‐FTICR‐MS approach was evaluated by using bovine serum albumin as a standard protein. We applied this approach to the protein identification of rat liver diacetyl‐reducing enzyme. It was demonstrated that this enzyme was correctly identified as 3‐α‐hydroxysteroid dehydrogenase by the alternating‐scan nano‐LC/IRMPD‐FTICR‐MS approach with accurate peptide mass fingerprinting and peptide sequence tagging. Copyright © 2003 John Wiley & Sons, Ltd.