Search of sequence databases with uninterpreted high-energy collision-induced dissociation spectra of peptides
Search of sequence databases with uninterpreted high-energy collision-induced dissociation spectra of peptides
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DOI：
10.1016/s1044-0305(96)00079-7
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发表时间：
1996-11-01
影响因子：
3.2
通讯作者：
Burlingame, AL
中科院分区：
文献类型：
--
作者：
Yates, JR;Eng, JK;Burlingame, AL
我们扩大了SEQUEST计算机算法的用途，允许将未解释的高能碰撞诱导的多肽解离光谱与数据库中的所有序列进行关联。SEQUEST现在允许在高能条件下观察到的额外碎片离子类型。我们分析了胰酶消化13种蛋白质后分离出的多肽的光谱。在OWL数据库的搜索中，SEQUEST将90%(18/20)的光谱的正确序列排在第一位，不受酶切割特异性或起源物种的限制。所有的假阳性都被计分系统标记出来。SEQUEST在数据库中搜索对应于前体离子质量+/-0.5u的序列。通过计算每个序列的碎片离子质量，并根据离子系列的连续性、总离子强度和离子的存在与测量的离子质量进行比较，对前500个候选离子进行初步排序。通过构建500个候选的模型谱并构建/执行与实际谱的互相关分析来进行最终排序。鉴于需要将不断增加的基因组序列信息与构成细胞分子结构的相应蛋白质组联系起来，串联质谱仪似乎注定要在大规模加速蛋白质鉴定方面发挥主导作用。(C)1996年美国质谱学学会
We have broadened the utility of the SEQUEST computer algorithm to permit correlation of uninterpreted high-energy collision-induced dissociation spectra of peptides with all sequences in a database. SEQUEST now allows for the additional fragment ion types observed under high-energy conditions. We analyzed spectra from peptides isolated following trypsin digestion of 13 proteins. SEQUEST ranked the correct sequence first for 90% (18/20) of the spectra in searches of the OWL database, without constraint by enzyme cleavage specificity or species of origin. All false-positives were flagged by the scoring system. SEQUEST searches databases for sequences that correspond to the precursor ion mass +/- 0.5 u. Preliminary ranking of the top 500 candidates is done by calculation of fragment ion masses for each sequence, and comparison to the measured ion masses on the basis of ion series continuity, summed ion intensity, and immonium ion presence. Final ranking is done by construction of model spectra for the 500 candidates and constructing/performing of a cross-correlation analysis with the actual spectrum. Given the need to relate mounting genome sequence information with corresponding suites of proteins that comprise the cellular molecular machinery, tandem mass spectrometry appears destined to play the leading role in accelerating protein identification on the large scale required. (C) 1996 American Society for Mass Spectrometry