Direct ionization of large proteins and protein complexes by desorption electrospray ionization-mass spectrometry.
Direct ionization of large proteins and protein complexes by desorption electrospray ionization-mass spectrometry.
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DOI：
10.1021/ac201390w
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发表时间：
2011-09-01
影响因子：
7.4
通讯作者：
Chen, Hao
中科院分区：
文献类型：
--
作者：
Ferguson, Carly N.;Benchaar, Sabrina A.;Miao, Zhixin;Loo, Joseph A.;Chen, Hao
解吸电喷雾电离质谱 (DESI-MS) 具有快速样品分析的优点，无需或很少进行样品预处理，但其对大生物分子的性能尚未得到证实。在这项研究中，液体样品 DESI（用于分析液体样品的 DESI 的扩展版本）被证明具有直接电离大型非共价蛋白质复合物（> 45 kDa）和蛋白质（高达 150 kDa）的能力。通过液体样品 DESI 从溶液中解吸的蛋白质复合离子（例如超氧化物歧化酶、烯醇酶和血红蛋白）被完整地测量，表明 DESI 保持弱非共价相互作用的能力。用增压试剂掺杂 DESI 喷雾溶剂导致蛋白质复合离子增加了多重电荷，而没有复合解离。模型蛋白细胞色素 c 的离子淌度测量表明，增压试剂有利于带较低电荷的蛋白离子形成更紧凑的构象。疏水性肽短杆菌肽 D 的液体样品 DESI 表明电离机制涉及液滴拾取混合过程。液体样品的测量显着扩展了 DESI-MS 的质量范围，允许分析高质量蛋白质，例如 150 kDa 免疫球蛋白 G (IgG)，因此代表了迄今为止 DESI 成功电离的最大蛋白质。
Desorption electrospray ionization-mass spectrometry (DESI-MS) has advantages for rapid sample analysis with little or no sample pretreatment, but performance for large biomolecules has not been demonstrated. In this study, liquid sample DESI, an extended version of DESI used for analysis of liquid sample, was shown to have capabilities for direct ionization of large noncovalent protein complexes (>45 kDa) and proteins (up to 150 kDa). Protein complex ions (e.g., superoxide dismutase, enolase, and hemoglobin) desorbed from solution by liquid sample DESI were measured intact, indicating the capability of DESI for preserving weak noncovalent interactions. Doping the DESI spray solvent with supercharging reagents resulted in protein complex ions having increased multiple charging without complex dissociation. Ion mobility measurements of model protein cytochrome c showed that the supercharging reagent favored the more compact conformation for the lower charged protein ions. Liquid sample DESI of hydrophobic peptide gramicidin D suggests that the ionization mechanism involves a droplet pick-up mixing process. Measurement of liquid samples significantly extends the mass range of DESI-MS, allowing the analysis of high-mass proteins such as 150 kDa immunoglobulin G (IgG), and thus represents the largest protein successfully ionized by DESI to date.
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影响因子：
7.4
作者：
Cody, RB;Laramée, JA;Durst, HD
通讯作者：
Durst, HD
影响因子：
4.9
作者：
Chen, H;Cotte-Rodríguez, I;Cooks, RG
通讯作者：
Cooks, RG
影响因子：
4.9
作者：
Chen, HW;Venter, A;Cooks, RG
通讯作者：
Cooks, RG
DOI：
10.1016/j.jasms.2009.09.014
发表时间：
2010-01
影响因子：
3.2
作者：
Lomeli SH;Peng IX;Yin S;Loo RR;Loo JA
通讯作者：
Loo JA
DOI：
10.1016/j.jasms.2008.11.013
发表时间：
2009-04-01
影响因子：
3.2
作者：
Lomeli, Shirley H.;Yin, Sheng;Loo, Joseph A.
通讯作者：
Loo, Joseph A.