Shevchenko A, Loboda A, Shevchenko A, Ens W, Standing K G
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Anal Chem. 2000 May 1;72(9):2132-41. doi: 10.1021/ac9913659.
A MALDI QqTOF mass spectrometer has been used to identify proteins separated by one-dimensional or two-dimensional gel electrophoresis at the femtomole level. The high mass resolution and the high mass accuracy of this instrument in both MS and MS/MS modes allow identification of a protein either by peptide mass fingerprinting of the protein digest or from tandem mass spectra acquired by collision-induced dissociation of individual peptide precursors. A peptide mass map of the digest and tandem mass spectra of multiple peptide precursor ions can be acquired from the same sample in the course of a single experiment. Database searching and acquisition of MS and MS/MS spectra can be combined in an interactive fashion, increasing the information value of the analytical data. The approach has demonstrated its usefulness in the comprehensive characterization of protein in-gel digests, in the dissection of complex protein mixtures, and in sequencing of a low molecular weight integral membrane protein. Proteins can be identified in all types of sequence databases, including an EST database. Thus, MALDI QqTOF mass spectrometry promises to have remarkable potential for advancing proteomic research.
基质辅助激光解吸电离串联飞行时间质谱仪已被用于在飞摩尔水平上鉴定通过一维或二维凝胶电泳分离的蛋白质。该仪器在质谱（MS）和串联质谱（MS/MS）模式下都具有高质量分辨率和高精度质量，这使得可以通过蛋白质消化产物的肽质量指纹图谱或通过单个肽前体的碰撞诱导解离获得的串联质谱来鉴定蛋白质。在单个实验过程中，可以从同一样品中获取消化产物的肽质量图谱和多个肽前体离子的串联质谱。数据库搜索以及MS和MS/MS谱图的采集可以以交互式方式结合起来，增加分析数据的信息价值。该方法已在蛋白质凝胶内消化产物的全面表征、复杂蛋白质混合物的剖析以及低分子量整合膜蛋白的测序中证明了其有用性。蛋白质可以在所有类型的序列数据库中进行鉴定，包括EST数据库。因此，基质辅助激光解吸电离串联飞行时间质谱有望在推进蛋白质组学研究方面具有显著潜力。
