期刊
EUROPEAN JOURNAL OF MASS SPECTROMETRY
卷 15, 期 6, 页码 739-746出版社
SAGE PUBLICATIONS LTD
DOI: 10.1255/ejms.1030
关键词
accessible surface area; calmodulin; hydrogen/deuterium exchange; MALDI; S100b; back-exchange
资金
- Barrow Neurological Foundation
Hydrogen/deuterium exchange (HDX) using matrix-assisted laser desorption ionization time-of-flight (MALDI-ToF) mass spectrometry is a sensitive, salt-tolerant and high-throughput method useful to probe protein conformation and molecular interactions. However, a drawback of the MALDI HDX technique is that sample preparation methods can typically result in higher levels of artificial deuterium in-exchange and/or hydrogen back-exchange just prior to or during mass analysis; this may impair data reproducibility and impede structural and kinetic data interpretation. White methods to minimize effects of back-exchange during protein analyte deposition on MALDI plates have been reported, this study presents a readily available, highly sensitive protein control set to facilitate rapid MALDI HDX protocol workup. The Ca2+-induced solvent accessible surface area (ASA) changes of calmodulin (CaM) and S100 proteins were employed to monitor and optimize HDX protocol efficiency. Under non-stringent room temperature conditions, the Ca2+-induced deuterium exchange of CaM, Delta Dca2+-apo, MH+ shifts -17 Da to -24 Da, white S100 Delta Dca2+-apo MH+ shifts +8 Da to +12 Da. By comparing the divergent CaM and S100 Ca2+-induced deuterium mass shift differences, HDX sample workup and MALDI plate spotting conditions can easily be monitored.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据