Future directions of structural mass spectrometry using hydroxyl radical footprinting.
Future directions of structural mass spectrometry using hydroxyl radical footprinting.
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DOI：
10.1002/jms.1808
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发表时间：
2010-12
影响因子：
2.3
通讯作者：
Chance, Mark R.
中科院分区：
文献类型：
--
作者：
Kiselar, Janna G.;Chance, Mark R.
关键词：
Hydroxyl radical protein footprinting coupled to mass spectrometry has been developed over the last decade and has matured to a powerful method for analyzing protein structure and dynamics. It has been successfully applied in the analysis of protein structure, protein folding, protein dynamics, and protein-protein and protein-DNA interactions. Using synchrotron radiolysis, exposures of proteins to a “white” x-ray beam for milliseconds provide sufficient oxidative modifications to surface amino acid side chains that can be easily detected and quantified by mass spectrometry. Thus, conformational changes in proteins or protein complexes can be examined using a time-resolved approach, which would be a valuable method for the study of macromolecular dynamics. In this review, we describe a new application of hydroxyl radical protein footprinting to probe the time evolution of the calcium-dependent conformational changes of gelsolin on the millisecond timescale. The data suggest a cooperative transition as multiple sites in different molecular sub-domains have similar rates of conformational change. These findings demonstrate that time-resolved protein footprinting is suitable for studies of protein dynamics that occur over periods ranging from milliseconds to seconds. In this review we also show how the structural resolution and sensitivity of the technology can be improved as well. The hydroxyl radical varies in its reactivity to different side chains by over two orders of magnitude, thus oxidation of amino acid side chains of lower reactivity are more rarely observed in such experiments. Here we demonstrate that selected reaction monitoring (SRM)-based method can be utilized for quantification of oxidized species, improving the signal to noise ratio. This expansion of the set of oxidized residues of lower reactivity will improve the overall structural resolution of the technique. This approach is also suggested as a basis for developing hypothesis driven structural mass spectrometry experiments.
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影响因子：
1.3
作者：
Novak, Petr;Giannakopulos, Anastassios E.
通讯作者：
Giannakopulos, Anastassios E.
影响因子：
7.4
作者：
Aye, TT;Low, TY;Sze, SK
通讯作者：
Sze, SK
DOI：
10.1073/pnas.1232301100
发表时间：
2003-06-10
影响因子：
11.1
作者：
Englander, JJ;Del Mar, C;Woods, VL
通讯作者：
Woods, VL
影响因子：
4.8
作者：
Ashish;Paine, Matthew S.;Krueger, Joanna K.
通讯作者：
Krueger, Joanna K.
DOI：
10.1016/j.str.2008.04.016
发表时间：
2008-08-06
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Bohon J;Jennings LD;Phillips CM;Licht S;Chance MR
通讯作者：
Chance MR