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Analysis of post‐translational modification and characterization of the domain structure of dynamin A from Dictyostelium discoideum
Author(s) -
Schlosser Andreas,
Klockow Boris,
Manstein Dietmar J.,
Lehmann Wolf D.
Publication year - 2003
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.438
Subject(s) - chemistry , dictyostelium discoideum , tandem mass spectrometry , mass spectrometry , covalent bond , protein mass spectrometry , gel electrophoresis , top down proteomics , chromatography , peptide , molecular mass , fast atom bombardment , electrospray ionization , biochemistry , organic chemistry , enzyme , gene
The post‐translational modifications of the 96 kDa protein dynamin A from Dictyostelium discoideum were analyzed using Q‐TOF mass spectrometry. The accurate molecular mass of the intact protein revealed a covalent modification causing an additional mass of 42 Da. The modification could be identified as N‐terminal acetylation by tandem mass spectrometry. Extracted ion chromatograms for the a 1 and b 1 ion of the tryptic T1 peptide were used to detect the acetylated peptide within 54 nanoelectrospray ionization tandem mass spectra. Owing to the accurate molecular mass of the intact protein, additional covalent modifications could be excluded. In addition to the covalent modification, the domain structure of dynamin A was determined by applying a combination of limited proteolysis, sodium dodecylsulfate polyacrylamide gel electrophoresis, automated tandem mass spectrometry and protein database searching. Copyright © 2003 John Wiley & Sons, Ltd.