Site-specific characterization of (D)-amino acid containing peptide epimers by ion mobility spectrometry.
Site-specific characterization of (D)-amino acid containing peptide epimers by ion mobility spectrometry.
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通过离子迁移率光谱法含有（D）氨基酸的（D） - 氨基酸的位点特异性表征。
DOI：
10.1021/ac4033824
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发表时间：
2014-03-18
影响因子：
7.4
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Jia, Chenxi;Lietz, Christopher B.;Yu, Qing;Li, Lingjun
传统上，通过观察合成多肽和天然多肽在生物活性和色谱保留时间上的差异，可以发现含有d-氨基酸的多肽(DAACP)候选分子。然而，很难确定d-氨基酸在DAACP候选分子中的确切位置。在此，我们开发了一种新的位点特异性策略，通过对质谱学(MS)产生的表观碎片离子进行离子迁移率(IMS)分析，快速准确地定位多肽中的d-氨基酸。简单地说，用在线反相液相色谱分离d/L-多肽同分异构体，然后用碰撞诱导解离(CID)和离子体质谱(IMS)分析。由d/L多肽同分异构体产生的表观碎片离子具有不同的构象，从而在IMS中表现出不同的迁移率。以表观碎片离子之间的到达时移作为d-氨基酸取代的定位标准。通过对不同分子大小的多肽同分异构体[d-Trp]-黑素细胞刺激素、[d-丙氨酸]-Deltorphin、[d-Phe]-Achatin-I以及含有全L氨基酸的对应多肽同分异构体的分析，证明了该策略的有效性。此外，从美国龙虾(Homarus Americanus)中分离到甲壳类高血糖激素(CHHS，8.5 kDa)，并结合基于MS的自下而上和自上而下的测序方法进行了鉴定。利用我们新的位点特异性策略获得的IMS数据定位了L异构化为d-Phe的位置在CHHS的N-末端的第三个残基上。总之，这项研究展示了一种利用IMS技术发现DAACPs的新方法，该方法具有定位d-氨基酸残基的能力。
Traditionally, the d-amino acid containing peptide (DAACP) candidate can be discovered by observing the differences of biological activity and chromatographic retention time between the synthetic peptides and naturally occurring peptides. However, it is difficult to determine the exact position of d-amino acid in the DAACP candidates. Herein, we developed a novel site-specific strategy to rapidly and precisely localize d-amino acids in peptides by ion mobility spectrometry (IMS) analysis of mass spectrometry (MS)-generated epimeric fragment ions. Briefly, the d/l-peptide epimers were separated by online reversed-phase liquid chromatography and fragmented by collision-induced dissociation (CID), followed by IMS analysis. The epimeric fragment ions resulting from d/l-peptide epimers exhibit conformational differences, thus showing different mobilities in IMS. The arrival time shift between the epimeric fragment ions was used as criteria to localize the d-amino acid substitution. The utility of this strategy was demonstrated by analysis of peptide epimers with different molecular sizes, [d-Trp]-melanocyte-stimulating hormone, [d-Ala]-deltorphin, [d-Phe]-achatin-I, and their counterparts that contain all-l amino acids. Furthermore, the crustacean hyperglycemia hormones (CHHs, 8.5 kDa) were isolated from the American lobster Homarus americanus and identified by integration of MS-based bottom-up and top-down sequencing approaches. The IMS data acquired using our novel site-specific strategy localized the site of isomerization of l- to d-Phe at the third residue of the CHHs from the N-terminus. Collectively, this study demonstrates a new method for discovery of DAACPs using IMS technique with the ability to localize d-amino acid residues.
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影响因子：
7.3
作者：
Grieco, P;Balse, PM;Hruby, VJ
通讯作者：
Hruby, VJ
影响因子：
7.4
作者：
Adams, CM;Zubarev, RA
通讯作者：
Zubarev, RA
影响因子：
2.7
作者：
Chung, J. Sook;Zmora, N.;Tsutsui, N.
通讯作者：
Tsutsui, N.
影响因子：
5.4
作者：
Ollivaux, Celine;Gallois, Dominique;Soyez, Daniel
通讯作者：
Soyez, Daniel
影响因子：
3.2
作者：
Serrano, L;Grousset, E;Spanings-Pierrot, C
通讯作者：
Spanings-Pierrot, C