Sulfate radical anion as a new reagent for fast photochemical oxidation of proteins.
Sulfate radical anion as a new reagent for fast photochemical oxidation of proteins.
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DOI：
10.1021/ac101760y
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发表时间：
2010-09-15
影响因子：
7.4
通讯作者：
Gross, Michael L.
中科院分区：
文献类型：
--
作者：
Gau, Brian C.;Chen, Hao;Zhang, Yun;Gross, Michael L.
重点是扩展蛋白质快速光化学氧化的原始设计，并引入SO4−·作为蛋白质足迹中的自由基反应物。FPOP是一种化学足迹方法，通过与自由基快照反应来确定蛋白质和蛋白质复合体的足迹。到目前为止使用的自由基是OH自由基，它提供了一种衡量天然蛋白质残基分解溶剂可及性的指标。我们表明，FPOP可以容纳其他试剂，增加了它的通用性。新的过硫酸盐FPOP系统是一种有效的、非特异性的和可调的足迹方法；需要减少3-5倍的过硫酸盐才能提供与OH自由基相同的全局修饰水平。尽管暴露在溶剂中的His和Tyr残基与SO_4−·的反应比与·OH的反应更强，但去肌红蛋白和钙调蛋白的氧化显示·OH探测的可及区域比SO_4−·小，可能组氨酸除外。His64是无肌红蛋白血红素结合袋中的轴向配体，被SO4−·相对于·OH大量上调。然而，这两种试剂自由基的修饰种类和残基选择性惊人地相似。因此，这些试剂的选择取决于自由基前驱体的物理性质，特别是膜的渗透性。
The focus is to expand the original design of fast photochemical oxidation of proteins (FPOP) and introduce SO4−•, generated by 248 nm homolysis of low mM levels of persulfate, as a radical reactant in protein footprinting. FPOP is a chemical footprinting approach to footprinting proteins and protein complexes by “snapshot” reaction with free radicals. The radical used until now is the OH radical, and it provides a measure of residue-resolved solvent accessibility of the native protein. We show that FPOP can accommodate other reagents, increasing its versatility. The new persulfate FPOP system is a potent, non-specific, and tunable footprinting method; 3–5 times less persulfate is needed to give the same global levels of modification as seen with OH radicals. Although solvent-exposed His and Tyr residues are more reactive with SO4−• than with •OH, oxidation of apomyoglobin and calmodulin shows that •OH probes smaller accessible areas than SO4−•, with the possible exception of histidine. His64, an axial ligand in the heme-binding pocket of apomyoglobin, is substantially up-labeled by SO4−• relative to •OH. Nevertheless, the kinds of modification and residue selectivity for both reagent radicals are strikingly similar. Thus, the choice of these reagents relies on the physical properties, particularly the membrane permeability, of the radical precursors.
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影响因子：
8
作者：
Craig, PO;Ureta, DB;Delfino, JM
通讯作者：
Delfino, JM
DOI：
10.1073/pnas.96.11.6020
发表时间：
1999-05-25
影响因子：
11.1
作者：
Fancy, DA;Kodadek, T
通讯作者：
Kodadek, T
影响因子：
15
作者：
CHATGILIALOGLU, C;LUNAZZI, L;PLACUCCI, G
通讯作者：
PLACUCCI, G
DOI：
10.1039/p29840000503
发表时间：
1984-01-01
期刊：
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 2
影响因子：
--
作者：
DAVIES, MJ;GILBERT, BC;NORMAN, ROC
通讯作者：
NORMAN, ROC
影响因子：
8
作者：
Gómez, GE;Cauerhff, A;Delfino, JM
通讯作者：
Delfino, JM