Shevchenko A, Sunyaev S, Loboda A, Shevchenko A, Bork P, Ens W, Standing K G
Peptide and Protein Group, European Molecular Biology Laboratory, Heidelberg, Germany.
Anal Chem. 2001 May 1;73(9):1917-26. doi: 10.1021/ac0013709.
MALDI-quadrupole time-of-flight mass spectrometry was applied to identify proteins from organisms whose genomes are still unknown. The identification was carried out by successively searching a sequence database-first with a peptide mass fingerprint, then with a packet of noninterpreted MS/MS spectra, and finally with peptide sequences obtained by automated interpretation of the MS/MS spectra. A "MS BLAST" homology searching protocol was developed to overcome specific limitations imposed by mass spectrometric data, such as the limited accuracy of de novo sequence predictions. This approach was tested in a small-scale proteomic project involving the identification of 15 bands of gel-separated proteins from the methylotrophic yeast Pichia pastoris, whose genome has not yet been sequenced and which is only distantly related to other fungi.
基质辅助激光解吸电离-四极杆飞行时间质谱法被用于鉴定来自基因组仍未知的生物体的蛋白质。鉴定过程是通过依次搜索序列数据库来进行的：首先使用肽质量指纹图谱，然后使用一组未解析的串联质谱（MS/MS）谱，最后使用通过对MS/MS谱进行自动解析获得的肽序列。开发了一种“MS BLAST”同源性搜索协议，以克服质谱数据所带来的特定限制，例如从头测序预测的准确性有限。这种方法在一个小规模蛋白质组学项目中进行了测试，该项目涉及鉴定来自甲基营养型酵母巴斯德毕赤酵母的15条凝胶分离蛋白条带，其基因组尚未测序，并且与其他真菌的亲缘关系较远。
