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Mapping the phosphorylation sites of proteins using on‐line immobilized metal affinity chromatography/capillary electrophoresis/electrospray ionization multiple stage tandem mass spectrometry
Author(s) -
Cao Ping,
Stults John T.
Publication year - 2000
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 - 10.1002/1097-0231(20000915)14:17<1600::aid-rcm68>3.0.co;2-v
Subject(s) - chemistry , chromatography , capillary electrophoresis , tandem mass spectrometry , electrospray ionization , mass spectrometry , capillary electrophoresis–mass spectrometry , protein mass spectrometry , sample preparation in mass spectrometry , electrospray
On‐line immobilized metal affinity chromatography/capillary electrophoresis/electrospray ionization‐mass spectrometry (IMAC/CE/ESI‐MS) offers selective preconcentration of phosphorylated peptides with identification of the phosphorylated amino acid(s). The preconcentration provides low concentration limits of detection and capillary electrophoresis separates the peptides. Recently, we reported a fast, simple, and sensitive on‐line IMAC/CE/ESI‐MS/MS method for the determination of phosphopeptides at low‐pmole levels. That work is expanded here by use of multiple stage tandem mass spectrometry (MS n , n = 2,3) to isolate and fragment target ions to provide more reliable assignments of phosphorylated residues. The application of IMAC/CE/ESI‐MS n is demonstrated by the analysis of tryptic digests of α‐ and β‐casein and in‐gel tryptic digests of β‐casein. Copyright © 2000 John Wiley & Sons, Ltd.