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Structural analysis of a highly acetylated protein using a curved‐field reflectron mass spectrometer
Author(s) -
Wang Dongxia,
Thompson Paul,
Cole Philip A.,
Cotter Robert J.
Publication year - 2005
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/pmic.200401167
Subject(s) - acetylation , mass spectrometry , lysine , tandem mass spectrometry , reflectron , chemistry , chromatography , protein mass spectrometry , isobaric labeling , trypsin , sample preparation in mass spectrometry , p300 cbp transcription factors , acetyltransferase , biochemistry , electrospray ionization , organic chemistry , time of flight mass spectrometry , ionization , amino acid , histone acetyltransferases , enzyme , ion , gene
Matrix‐assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry (MS/MS) were used to determine the multiple acetylation sites in the histone acetyltransferase (HAT): p300‐HAT. Partial cleavage of the peptides containing acetylated lysine residues by trypsin provided a set of nested sequences that enabled us to determine that multiple acetylation occurs on the same molecule. At the same time, cleavages resulting in a terminal unacetylated lysine suggested that not all of these sites are fully modified. Using MS and MS/MS, we were able to characterize both the unmodified and acetylated tryptic peptides covering more than 82% of the protein.