New reagents for increasing ESI multiple charging of proteins and protein complexes.
New reagents for increasing ESI multiple charging of proteins and protein complexes.
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DOI：
10.1016/j.jasms.2009.09.014
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发表时间：
2010-01
影响因子：
3.2
通讯作者：
Loo JA
中科院分区：
文献类型：
--
作者：
Lomeli SH;Peng IX;Yin S;Loo RR;Loo JA
之前已经证明了间硝基苯甲醇（m-NBA）的加入(Lomeli et al.， J. Am.；Soc。质谱。2009,20,593-596)在电喷雾电离（ESI）质谱中增强天然蛋白和非共价蛋白复合物的多次充电。人们还发现了一些新的试剂，可以使蛋白质从非变性溶液中“增压”；这些试剂中的一些被证明比m-NBA更有效地增加正电荷。使用肌红蛋白-原卟啉IX（血红素）配合物，发现以下试剂可以增加ESI电荷：苯甲醇、间硝基苯乙酮、间硝基苯腈、o-NBA、间nba、对- nba、间硝基苯基乙醇、亚砜（四亚甲基砜）和间（三氟甲基）苯甲醇。基于平均电荷状态，在水溶液中，亚砜对肌红蛋白的电荷增加（61%）大于m-NBA（21%）。促进较高ESI电荷的试剂具有较低的固相碱度和相对较低的气相碱度，并且比水挥发性更小。一些活性试剂的质谱的另一个特征是加合物存在于较高的电荷状态，这表明蛋白质获得额外电荷的机制除了表面张力和蛋白质变性等其他因素外，还包括与试剂的直接相互作用。
The addition of m-nitrobenzyl alcohol (m-NBA) was shown previously (Lomeli et al., J. Am. Soc. Mass Spectrom. 2009, 20, 593–596) to enhance multiple charging of native proteins and noncovalent protein complexes in electrospray ionization (ESI) mass spectra. Additional new reagents have been found to “supercharge” proteins from nondenaturing solutions; several of these reagents are shown to be more effective than m-NBA for increasing positive charging. Using the myoglobin protein-protoporphyrin IX (heme) complex, the following reagents were shown to increase ESI charging: benzyl alcohol, m-nitroacetophenone, m-nitrobenzonitrile, o-NBA, m-NBA, p-NBA, m-nitrophenyl ethanol, sulfolane (tetramethylene sulfone), and m-(trifluoromethyl)-benzyl alcohol. Based on average charge state, sulfolane displayed a greater charge increase (61%) than m-NBA (21%) for myoglobin in aqueous solutions. The reagents that promote higher ESI charging appear to have low solution-phase basicities and relatively low gas-phase basicities, and are less volatile than water. Another feature of mass spectra from some of the active reagents is that adducts are present on higher charge states, suggesting that a mechanism by which proteins acquire additional charge involves direct interaction with the reagent, in addition to other factors such as surface tension and protein denaturation.
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DOI：
10.1016/0168-1176(87)87041-6
发表时间：
1987-09-24
期刊：
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY AND ION PROCESSES
影响因子：
--
作者：
KARAS, M;BACHMANN, D;HILLENKAMP, F
通讯作者：
HILLENKAMP, F
影响因子：
5.1
作者：
PETTIT, GR;HOLZAPFEL, CW;WILLIAMS, P
通讯作者：
WILLIAMS, P
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.
影响因子：
7.4
作者：
Li, Y;Cole, RB
通讯作者：
Cole, RB
影响因子：
56.9
作者：
LOO, JA;EDMONDS, CG;SMITH, RD
通讯作者：
SMITH, RD