The influence of a C-terminal basic residue on peptide fragmentation pathways
The influence of a C-terminal basic residue on peptide fragmentation pathways
复制标题
DOI：
10.1016/j.ijms.2012.02.017
复制
发表时间：
2012-04-15
影响因子：
1.8
通讯作者：
Gaskell, Simon J.
中科院分区：
文献类型：
--
作者：
Chawner, Ross;Eyers, Claire E.;Gaskell, Simon J.
典型的“自下而上”蛋白质组学工作流程使用串联质谱数据来推断产物离子序列，从而推断它们所来源的蛋白质的身份。这种分析通常在用内切蛋白酶胰蛋白酶或Lys-C进行蛋白水解后进行;因此产生的肽终止于碱性残基精氨酸或赖氨酸。使用外肽酶羧肽酶B除去这些C-末端碱性残基，产生肽，其通过串联MS的分析产生基本上不同的片段化性质的证据。用碰撞诱导解离和电子转移解离研究了用羧肽酶B处理之前和之后肽离子的分解。二次酶处理后的性质的变化主要归因于去除强碱性位点，从而对保留电荷的倾向和碎片离子的稳定性产生影响。这些数据表明，在蛋白质组分析中，胰蛋白酶/Lys-C肽的MS/MS在随后的羧肽酶B处理和不处理的情况下具有互补价值。(C)2012爱思唯尔有限公司版权所有。
A typical 'bottom up' proteomic workflow uses tandem mass spectrometric data to infer product ion sequence and hence identity of the protein from which they derive. Such analysis is typically performed following proteolysis with the endoproteases trypsin or Lys-C; peptides produced therefore terminate in the basic residues arginine or lysine. Removal of these C-terminal basic residues using the exopeptidase, carboxypeptidase B, generates peptides whose analysis by tandem MS yields evidence of substantially different fragmentation properties. The decompositions of peptide ions both prior to and following treatment with carboxypeptidase B have been examined using collision-induced dissociation and electron transfer dissociation. Changes in properties following secondary enzyme treatment are attributed primarily to removal of a strongly basic site, with a consequent effect both on the propensity to retain charge and the stability of the fragment ions. The data suggest a complementary value in proteome analyses for MS/MS of tryptic/Lys-C peptides with and without subsequent carboxypeptidase B treatment. (C) 2012 Elsevier B.V. All rights reserved.