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Identification of p21 Cip1 binding proteins by gel electrophoresis and capillary liquid chromatography microelectrospray tandem mass spectrometry
Author(s) -
Carrascal Montserrat,
Carujo Sonia,
Bachs Oriol,
Abian Joaquin
Publication year - 2002
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200204)2:4<455::aid-prot455>3.0.co;2-e
Subject(s) - chromatography , capillary electrophoresis , mass spectrometry , chemistry , tandem mass spectrometry , tandem mass tag , liquid chromatography–mass spectrometry , two dimensional gel electrophoresis , proteomics , top down proteomics , protein mass spectrometry , capillary electrophoresis–mass spectrometry , tandem , gel electrophoresis , quantitative proteomics , biochemistry , electrospray ionization , materials science , composite material , gene
Proteins bound to a glutathione‐ S ‐transferase‐p21 Cip1 affinity column were separated by one‐dimensional sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and identified using tandem mass spectrometry. Capillary liquid chromatography coupled to microelectrospray tandem mass spectrometry (capLC‐νESI MS/MS) in an ion trap allowed identification of the proteins present in the gel bands. Of eleven bands analyzed, fifty‐three proteins were identified. More than one hundred tryptic peptides were detected on‐line, automatically fragmented and used for protein characterization in databases. Samples were also analyzed by off‐line nanospray and matrix‐assisted laser desorption/ionization mass spectrometry. CapLC‐νESI MS/MS was the most efficient technique for the analysis of these protein mixtures.