Hydrogen exchange electrospray ionization mass spectrometry studies of structural features of proteins and protein/protein interactions
Hydrogen exchange electrospray ionization mass spectrometry studies of structural features of proteins and protein/protein interactions
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DOI：
10.1006/abio.1998.3000
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发表时间：
1999-02-15
影响因子：
2.9
通讯作者：
Ehring, H
中科院分区：
文献类型：
--
作者：
Ehring, H
位于蛋白质/蛋白质复合物中肽主链上的酰胺氢进行氢/氘交换的速率高度依赖于酰胺基团是否参与结合。在这里，提出了一种新的质谱方法，其中利用这种效应来表征蛋白质/配体结合位点。获得的信息是蛋白质内的哪个区域参与结合。该方法包括受体和配体蛋白酰胺质子的氢/氘交换、结合和反向交换。在此过程之后，那些参与蛋白质结合的主链酰胺基团受到保护，免于反向交换，因此仍然被氘化。然后通过消化蛋白水解、快速微孔高效液相色谱分离和电喷雾电离质谱法来鉴定这些区域。该方法已应用于胰岛素样生长因子I（IGF-I）的结构特征以及胰岛素样生长因子I与IGF-I结合蛋白1的相互作用的研究。数据表明，该方法可以提供IGF-1疏水核心的位置以及主要参与与IGF-I结合蛋白1结合的两个区域的信息。数据与其他方法获得的结果一致。一项实验所需的样品量在亚纳摩尔范围内。 (C) 1999 年学术出版社。
The rate at which amide hydrogens located at the peptide backbone in protein/protein complexes undergo hydrogen/deuterium exchange is highly dependent on whether the amide groups participate in binding. Here, a new mass spectrometric method is presented in which this effect is utilized for the characterization of protein/ligand binding sites. The information obtained is which region within the protein participates in binding. The method includes hydrogen/deuterium exchange of receptor and ligand protein amide protons, binding, and back exchange. After this procedure those backbone amide groups that participate in protein binding are protected from back exchange and therefore still deuterated. These regions were then identified by peptic proteolysis, fast microbore high-performance liquid chromatography separation, and electrospray ionization mass spectrometry. The approach has been applied to the investigation of structural features of insulin-like growth factor I (IGF-I) and the interaction of insulin-like growth factor I with IGF-I binding protein 1. The data show that the approach can provide information on the location of the hydrophobic core of IGF-1 and on two regions that are mainly involved in binding to IGF-I binding protein 1. The data are consistent with results obtained with other approaches. The amount of sample required for one experiment is in the sub-nanomolar range. (C) 1999 Academic Press.