Schlosser Andreas, Klockow Boris, Manstein Dietmar J, Lehmann Wolf D
Central Spectroscopy, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.
J Mass Spectrom. 2003 Mar;38(3):277-82. doi: 10.1002/jms.438.
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.
利用Q-TOF质谱分析法对盘基网柄菌中96 kDa的发动蛋白A的翻译后修饰进行了分析。完整蛋白质的精确分子量显示存在一种共价修饰，导致分子量增加了42 Da。通过串联质谱法可将该修饰鉴定为N端乙酰化。在54个纳升电喷雾电离串联质谱图中，使用胰蛋白酶T1肽的a(1)和b(1)离子的提取离子色谱图来检测乙酰化肽段。由于完整蛋白质的精确分子量，可排除其他共价修饰。除了共价修饰外，还通过结合有限蛋白酶解、十二烷基硫酸钠聚丙烯酰胺凝胶电泳、自动串联质谱法和蛋白质数据库搜索来确定发动蛋白A的结构域结构。
