High-throughput bioconjugation for enhanced 193 nm photodissociation via droplet phase initiated ion/ion chemistry using a front-end dual spray reactor.
High-throughput bioconjugation for enhanced 193 nm photodissociation via droplet phase initiated ion/ion chemistry using a front-end dual spray reactor.
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DOI：
10.1021/acs.analchem.5b02242
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发表时间：
2015-09-15
影响因子：
7.4
通讯作者：
Brodbelt JS
中科院分区：
文献类型：
--
作者：
Cotham VC;Shaw JB;Brodbelt JS
利用线性离子阱（LIT）质谱仪前端的双电喷雾反应器，实现了芳香族标记肽的快速在线化学衍生化，增强了193 nm紫外光解离（UVPD）。该反应器促进质子化肽与第二群显色4-甲酰基-1，3-苯二磺酸（FBDSA）阴离子的交叉，以促进在大气压下实时形成离子/离子络合物。离子/离子中间体的随后碰撞活化导致经由肽阳离子中的伯胺与FBDSA阴离子的醛部分之间的反应生成的席夫碱形成。利用193 nm UVPD作为MS 3工作流程中的后续活化步骤，可获得相对于常规碰撞诱导解离（CID）更多的一级序列信息。此外，席夫碱修饰的肽与其未修饰的对应物相比，表现出平均20%的UVPD效率增加。由于与双喷雾反应器实现的共价标记的效率，我们证明，这种策略可以集成到一个高通量的LC-MSn工作流程，用于快速衍生化的肽混合物。
Fast on-line chemical derivatization of peptides with an aromatic label for enhanced 193 nm ultraviolet photodissociation (UVPD) is demonstrated using a dual electrospray reactor implemented on the front-end of a linear ion trap (LIT) mass spectrometer. The reactor facilitates the intersection of protonated peptides with a second population of chromogenic 4-formyl-1,3-benzenedisulfonic acid (FBDSA) anions to promote real-time formation of ion/ion complexes at atmospheric pressure. Subsequent collisional activation of the ion/ion intermediate results in Schiff base formation generated via reaction between a primary amine in the peptide cation and the aldehyde moiety of the FBDSA anion. Utilizing 193 nm UVPD as the subsequent activation step in the MS3 workflow results in acquisition of greater primary sequence information relative to conventional collision induced dissociation (CID). Furthermore, Schiff base modified peptides exhibit on average a 20% increase in UVPD efficiency compared to their unmodified counterparts. Due to the efficiency of covalent labeling achieved with the dual spray reactor, we demonstrate that this strategy can be integrated into a high-throughput LC-MSn workflow for rapid derivatization of peptide mixtures.