Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time of flight mass spectrometry and BLAST homology searching
Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time of flight mass spectrometry and BLAST homology searching
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DOI：
10.1021/ac0013709
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发表时间：
2001-05-01
影响因子：
7.4
通讯作者：
Standing, KG
中科院分区：
文献类型：
--
作者：
Shevchenko, A;Sunyaev, S;Standing, KG
MALDI-四极杆飞行时间质谱仪被用于从基因组仍未知的生物体中鉴定蛋白质。鉴定是通过连续搜索序列数据库进行的-首先使用肽质量指纹图谱，然后使用一包未解释的MS/MS谱，最后使用通过自动解释MS/MS谱获得的肽序列。“MS BLAST”同源搜索协议NAS的开发是为了克服质谱学数据施加的特定限制，例如从头序列预测的有限准确性。这种方法在一个小规模的蛋白质组项目中得到了测试，该项目涉及从甲醇营养酵母巴斯德毕赤酵母中鉴定15条凝胶分离蛋白，其基因组尚未测序，而且与其他真菌只有遥远的亲缘关系。
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 Nas 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.