title: Partial cooperative unfolding in proteins as observed by hydrogen exchange mass spectrometry.
abstract: Many proteins do not exist in a single rigid conformation. Protein motions, or dynamics, exist and in many cases are important for protein function. The analysis of protein dynamics relies on biophysical techniques that can distinguish simultaneously existing populations of molecules and their rates of interconversion. Hydrogen exchange (HX) detected by mass spectrometry (MS) is contributing to our understanding of protein motions by revealing unfolding and dynamics on a wide timescale, ranging from seconds to hours to days. In this review we discuss HX MS-based analyses of protein dynamics, using our studies of multi-domain kinases as examples. Using HX MS, we have successfully probed protein dynamics and unfolding in the isolated SH3, SH2 and kinase domains of the c-Src and Abl kinase families, as well as the role of inter- and intra-molecular interactions in the global control of kinase function. Coupled with high-resolution structural information, HX MS has proved to be a powerful and versatile tool for the analysis of the conformational dynamics in these kinase systems, and has provided fresh insight regarding the regulatory control of these important signaling proteins. HX MS studies of dynamics are applicable not only to the proteins we illustrate here, but to a very wide range of proteins and protein systems, and should play a role in both classification of and greater understanding of the prevalence of protein motion.
authors.affiliation: Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115 USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Engen
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Wales
authors.forename: Shugui
authors.initials: S
authors.lastname: Chen
authors.forename: Elaine M
authors.initials: EM
authors.lastname: Marzluff
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Hassell
authors.forename: David D
authors.initials: DD
authors.lastname: Weis
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Smithgall
keywords: Abl; HDX; Hck; Lck; SH2 domain; SH3 domain; Src-family kinase; deuterium; protein dynamics, flexibility