Increasing: Charge While Preserving Noncovalent Protein Complexes for ESI-MS
Increasing: Charge While Preserving Noncovalent Protein Complexes for ESI-MS
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DOI：
10.1016/j.jasms.2008.11.013
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发表时间：
2009-04-01
影响因子：
3.2
通讯作者：
Loo, Joseph A.
中科院分区：
文献类型：
--
作者：
Lomeli, Shirley H.;Yin, Sheng;Loo, Joseph A.
天然蛋白质和非共价蛋白质复合物的多个充电增加，观察到电喷雾电离（EST）质谱从非变性蛋白质溶液中含有高达1%（体积/体积）间硝基苄醇（m-NBA）。电荷增加的范围从690 kDa α（7）β（7）β（7）α（7）20 S蛋白酶体复合物的8%到锌结合的29 kDa碳酸酐酶-II蛋白的48%额外电荷。加入m-NBA后，未观察到非共价结合的配体/亚基的解离。尚不清楚增强的充电是否与Iavarone和威廉姆斯的“增压”模型中提出的改变的表面张力有关（Iavarone，A. T.;威廉姆斯，E. R. J. Am. 2003，125，2319-2327）。然而，更多的高度带电的非共价蛋白质复合物具有实用性，稍微放松的质量-电荷（m/z）的质谱仪检测的要求，并将有效地提高串联质谱研究的蛋白质复合物的效率。（J Am Soc Mass Spectrom 2009，20，593-596）（C）2009由Elsevier Inc.美国质谱学会American Society for Mass Spectrometry
Increased multiple charging of native proteins and noncovalent protein complexes is observed in electrospray ionization (EST) mass spectra obtained from nondenaturing protein solutions containing up to 1% (vol/vol) m-nitrobenzyl alcohol (m-NBA). The increases in charge ranged from 8% for the 690 kDa alpha(7)beta(7)beta(7)alpha(7) 20S proteasome complex to 48% additional charge for the zinc-bound 29 kDa carbonic anhydrase-II protein. No dissociation of the noncovalently bound ligands/subunits was observed upon the addition of m-NBA. It is not clear if the enhanced charging is related to altered surface tension as proposed in the "supercharging" model of Iavarone and Williams (Iavarone, A. T.; Williams, E. R. J. Am. Chem. Soc. 2003, 125, 2319-2327). However, more highly charged noncovalent protein complexes have utility in relaxing slightly the mass-to-charge (m/z) requirements of the mass spectrometer for detection and will be effective for enhancing the efficiency for tandem mass spectrometry studies of protein complexes. (J Am Soc Mass Spectrom 2009, 20, 593-596) (C) 2009 Published by Elsevier Inc. on behalf of American Society for Mass Spectrometry