Controlling deuterium isotope effects in comparative proteomics.
Controlling deuterium isotope effects in comparative proteomics.
复制标题
DOI：
10.1021/ac025614w
复制
发表时间：
2002-07
影响因子：
7.4
通讯作者：
Roujian Zhang;Cathy S Sioma;Robert A. Thompson;L. Xiong;F. Regnier
中科院分区：
文献类型：
--
作者：
Roujian Zhang;Cathy S Sioma;Robert A. Thompson;L. Xiong;F. Regnier
本文主要研究了在反相色谱中影响重同位素编码肽段分离的结构特征。这是通过使用标记的编码剂来实现的，所述标记的编码剂在结构、氘原子的数量、编码剂中氘的位置以及试剂靶向的官能团方面不同。检查了六种编码剂。所研究的编码剂的氘代形式包括琥珀酸酐-2H4、乙酸2，5-二氧代吡咯烷-1-基酯-2H3、丙酸2，5-二氧代吡咯烷-1-基酯-2H5、戊酸2，5-二氧代吡咯烷-1-基酯-2H9、[3-（2，5-二氧代吡咯烷-1-基氧基羰基）-丙基]-三甲基氯化铵-2H9和商业ICAT-2 H8试剂。结果发现，这些标记剂在它们对色谱同位素效应的绝对和相对贡献方面变化很大。通过将衍生肽中氘原子数的分辨率归一化来评价相对效应。单一的，最主要的影响是氘原子相对于编码剂中的亲水性官能团的位置。得出的结论是，氘原子与反相色谱（RPC）柱的固定相相互作用并影响分离度的可能性通过将其放置在亲水基团附近而大大降低，如疏溶剂理论所解释的。但是肽的大小和编码剂的大小也被认为与同位素效应的大小呈负相关。这种效应被解释为是由于编码剂相对于编码剂-肽缀合物的相对大小。
This paper focuses on identifying structural features responsible for resolution of heavy isotope coded peptides during reversed-phase chromatography. This was achieved by using labeled coding agents that varied in structure, number of deuterium atoms, placement of deuterium in the coding agent, and the functional group targeted by the reagent. Six coding agents were examined. Deuterated versions of the coding agents studied included succinic anhydride-2H4, acetic acid 2,5-dioxopyrrolidin-1-yl ester-2H3, propionic acid 2,5-dioxopyrrolidin-1-yl ester-2H5, pentanoic acid 2,5-dioxopyrrolidin-1-yl ester-2H9, [3-(2,5-dioxopyrrolidin-1-yloxycarbonyl)-propyl]-trimethylammonium chloride-2H9, and the commercial ICAT-2H8 reagent. It was found that these labeling agents vary widely in both their absolute and relative contribution to the chromatographic isotope effect. Relative effects were evaluated by normalizing resolution for the number of deuterium atoms in the derivatized peptide. The single, most dominant effect was the placement of deuterium atoms relative to hydrophilic functional groups in the coding agent. It was concluded that the probability of a deuterium atom interacting with the stationary phase of a reversed-phase chromatography (RPC) column and impacting resolution is greatly diminished by placing it adjacent to a hydrophilic group, as explained by solvophobic theory. But peptide size and coding agent size were also seen to correlate inversely with the magnitude of the isotope effect. This effect was explained as being due to the relative size of the coding agent versus that of the coding agent-peptide conjugate.