The effect of phosphorylation on the electron capture dissociation of peptide ions.
The effect of phosphorylation on the electron capture dissociation of peptide ions.
复制标题
DOI：
10.1016/j.jasms.2008.05.015
复制
发表时间：
2008-09
影响因子：
3.2
通讯作者：
Cooper HJ
中科院分区：
文献类型：
--
作者：
Creese AJ;Cooper HJ
研究了磷酸化位点和频率对肽离子电子捕获解离的影响。在一定范围的ECD阴极电位下，测定了一组合成肽（APLSFRGSLPKSYVK;一种未修饰的、三种单磷酸化的、三种双磷酸化的和一种三磷酸化的）、两种胰蛋白酶磷酸肽（YKVPQLEIVPNpSAEER，α-酪蛋白和YKVPSEEQQQTEDELQDK，β-酪蛋白）及其未修饰对应物的ECD。结果表明，对于双电荷的前体离子，磷酸化的存在对ECD序列覆盖率具有有害影响。观察到的片段化模式表明，对于具有多个碱性残基的肽，磷酸基团以其去质子化形式存在，并与质子化氨基酸侧链形成盐桥。观察到的酸性胰蛋白酶肽的碎片表明存在非共价相互作用，这是扰动磷酸化。增加ECD电子能量显著改善序列覆盖。或者，可通过对三价带电前体离子进行ECD来实现改进的序列覆盖。这一发现对于理解磷酸肽的气相裂解具有重要意义。由于去质子化的磷酸基团和质子化的赖氨酸/精氨酸侧链之间的盐桥形成，磷酸化对双电荷碱性肽的ECD具有有害影响。
The effect of site and frequency of phosphorylation on the electron capture dissociation of peptide ions has been investigated. The ECD of a suite of synthetic peptides (APLSFRGSLPKSYVK; one unmodified, three singly-phosphorylated, three-doubly phosphorylated, and one triply-phosphorylated); two tryptic phosphopeptides (YKVPQLEIVPNpSAEER, α-casein and FQpSEEQQQTEDELQDK, β-casein) and their unmodified counterparts, were determined over a range of ECD cathode potentials. The results show that, for doubly-charged precursor ions, the presence of phosphorylation has a deleterious effect on ECD sequence coverage. The fragmentation patterns observed suggest that for peptides with multiple basic residues, the phospho-groups exist in their deprotonated form and form salt-bridges with protonated amino acid side chains. The fragmentation observed for the acidic tryptic peptides suggested the presence of noncovalent interactions, which were perturbed on phosphorylation. Increasing the ECD electron energy significantly improves sequence coverage. Alternatively, improved sequence coverage can be achieved by performing ECD on triply-charged precursor ions. The findings are important for the understanding of gas-phase fragmentation of phosphopeptides. Phosphorylation has a deleterious effect on the ECD of doubly-charged basic peptides due to salt bridge formation between deprotonated phospho-groups and protonated lysine/arginine side chains.
登录
查看更多内容
影响因子：
7
作者：
Larsen, MR;Thingholm, TE;Jorgensen, TJD
通讯作者：
Jorgensen, TJD
影响因子：
4.4
作者：
Cooper, HJ;Akbarzadeh, S;Zeller, M
通讯作者：
Zeller, M
DOI：
10.1016/s1044-0305(02)00707-9
发表时间：
2002-12-01
影响因子：
3.2
作者：
Guan, ZQ
通讯作者：
Guan, ZQ
影响因子：
4.2
作者：
Kjeldsen, Frank;Savitski, Mikhail M.;Zubarev, Roman A.
通讯作者：
Zubarev, Roman A.
影响因子：
7.4
作者：
Mirgorodskaya, E;Roepstorff, P;Zubarev, RA
通讯作者：
Zubarev, RA