Protein structure and dynamics studied by mass spectrometry: H/D exchange, hydroxyl radical labeling, and related approaches

✍ Scribed by Lars Konermann; Xin Tong; Yan Pan

• Publisher: John Wiley and Sons
• Year: 2008
• Tongue: English
• Weight: 296 KB
• Volume: 43
• Category: Article
• ISSN: 1076-5174
• DOI: 10.1002/jms.1435

✦ Synopsis

Abstract

Mass spectrometry (MS) plays a central role in studies on protein structure and dynamics. This review highlights some of the recent developments in this area, with focus on applications involving the use of electrospray ionization (ESI) MS. Although this technique involves the transformation of analytes into highly nonphysiological species (desolvated gas‐phase ions in the vacuum), ESI–MS can provide detailed insights into the solution‐phase behavior of proteins. Notably, the ionization process itself occurs in a structurally sensitive manner. An increased degree of solution‐phase unfolding is correlated with a higher level of protonation. Also, ESI allows the transfer of intact noncovalent complexes into the gas phase, thereby yielding information on binding partners, stoichiometries, and even affinities. A particular focus of this article is the use of hydrogen/deuterium exchange (HDX) methods and hydroxyl radical (·OH) labeling for monitoring dynamic and structural aspect of solution‐phase proteins. Conceptual similarities and differences between the two methods are discussed. We describe a simple method for the computational simulation of protein HDX patterns, a tool that can be helpful for the interpretation of isotope exchange data recorded under mixed EX1/EX2 conditions. Important aspects of ·OH labeling include a striking dependence on protein concentration, and the tendency of commonly used solvent additives to act as highly effective radical scavengers. If not properly controlled, both of these factors may lead to experimental artifacts. Copyright © 2008 John Wiley & Sons, Ltd.