Automated data reduction for hydrogen/deuterium exchange experiments, enabled by high-resolution Fourier transform ion cyclotron resonance mass spectrometry.
Automated data reduction for hydrogen/deuterium exchange experiments, enabled by high-resolution Fourier transform ion cyclotron resonance mass spectrometry.
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DOI：
10.1016/j.jasms.2009.12.016
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发表时间：
2010-04
影响因子：
3.2
通讯作者：
Marshall AG
中科院分区：
文献类型：
--
作者：
Kazazic S;Zhang HM;Schaub TM;Emmett MR;Hendrickson CL;Blakney GT;Marshall AG
氢/氢交换后蛋白质降解片段多肽的质量分析提供了溶液相蛋白质及其络合物的溶剂可及性/氢键(从而构象)的一般度量。在这种质量分析中的主要问题是，在目标多肽、自溶产物和其他干扰物种的同位素分布存在实质性重叠的情况下，可靠和快速地将质谱峰指定为每个片段多肽的正确的电荷状态和氢化程度。在这里，我们展示了在足够高的质量分辨率(m/ΔM50%≥100,000)下，有可能解决足够多的重叠，使得基于每个肽的实际元素组成的自动数据简化成为可能，而不需要去卷积同位素分布。我们展示了基于溶液相肌红蛋白H/D交换的电喷雾电离FT-ICR质谱学，从H/D交换实验中自动、快速、可靠地指定多肽质量。与以前证明的用于此类实验的自动数据采集相结合，本数据简化算法增强了基于高分辨率质谱学的高分辨率质谱学分析溶液相蛋白质的H/D交换的自动化(从而扩展了通用性和适用性)。
Mass analysis of proteolytic fragment peptides following hydrogen/deuterium exchange offers a general measure of solvent accessibility/hydrogen bonding (and thus conformation) of solution-phase proteins and their complexes. The primary problem in such mass analyses is reliable and rapid assignment of mass spectral peaks to the correct charge state and degree of deuteration of each fragment peptide, in the presence of substantial overlap between isotopic distributions of target peptides, autolysis products, and other interferant species. Here, we show that at sufficiently high mass resolving power (m/Δm50% ≥ 100,000), it becomes possible to resolve enough of those overlaps so that automated data reduction becomes possible, based on the actual elemental composition of each peptide without the need to deconvolve isotopic distributions. We demonstrate automated, rapid, reliable assignment of peptide masses from H/D exchange experiments, based on electrospray ionization FT-ICR mass spectra from H/D exchange of solution-phase myoglobin. Combined with previously demonstrated automated data acquisition for such experiments, the present data reduction algorithm enhances automation (and thus expands generality and applicability) for high-resolution mass spectrometry- based analysis of H/D exchange of solution-phase proteins.
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影响因子：
8
作者：
Hotchko, M;Anand, GS;Ten Eyck, LF
通讯作者：
Ten Eyck, LF
DOI：
10.1016/j.jasms.2006.06.006
发表时间：
2006-11-01
影响因子：
3.2
作者：
Englander, S. Walter
通讯作者：
Englander, S. Walter
DOI：
10.1016/s1044-0305(02)00384-7
发表时间：
2002-06-01
影响因子：
3.2
作者：
Schwartz, JC;Senko, MW;Syka, JEP
通讯作者：
Syka, JEP
DOI：
10.1073/pnas.0812413106
发表时间：
2009-02-03
影响因子：
11.1
作者：
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通讯作者：
Demetri, George D.
影响因子：
3
作者：
Pascal BD;Chalmers MJ;Busby SA;Mader CC;Southern MR;Tsinoremas NF;Griffin PR
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