title: Quantitative measurement of exchange dynamics in proteins via (13)C relaxation dispersion of (13)CHD2-labeled samples.
abstract: Methyl groups have emerged as powerful probes of protein dynamics with timescales from picoseconds to seconds. Typically, studies involving high molecular weight complexes exploit (13)CH3- or (13)CHD2-labeling in otherwise highly deuterated proteins. The (13)CHD2 label offers the unique advantage of providing (13)C, (1)H and (2)H spin probes, however a disadvantage has been the lack of an experiment to record (13)C Carr-Purcell-Meiboom-Gill relaxation dispersion that monitors millisecond time-scale dynamics, implicated in a wide range of biological processes. Herein we develop an experiment that eliminates artifacts that would normally result from the scalar coupling between (13)C and (2)H spins that has limited applications in the past. The utility of the approach is established with a number of applications, including measurement of ms dynamics of a disease mutant of a 320 kDa p97 complex.
authors.affiliation: Departments of Molecular Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada.
authors.forename: Enrico
authors.initials: E
authors.lastname: Rennella
authors.affiliation: Departments of Molecular Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada.
authors.forename: Anne K
authors.initials: AK
authors.lastname: Schuetz
authors.affiliation: Departments of Molecular Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada. kay@pound.med.utoronto.ca.|Program in Molecular Structure and Function, Hospital for Sick Children, 555 University Avenue, Toronto, ON, M5G 1X8, Canada. kay@pound.med.utoronto.ca.
authors.forename: Lewis E
authors.initials: LE
authors.lastname: Kay
mesh_terms: D066241:Carbon-13 Magnetic Resonance Spectroscopy / Q000379:methods; D003903:Deuterium; D056004:Molecular Dynamics Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy
chemical_list: D011506:Proteins; D003903:Deuterium
keywords: 13CHD2; CPMG; Conformational dynamics; Methyl labeling; Proteins; Relaxation dispersion