Detection of chemical exchange in methyl groups of macromolecules
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The zero- and double-quantum methyl TROSY Hahn-echo and the methyl 1H–1H dipole–dipole cross-correlation nuclear magnetic resonance experiments enable estimation of multiple quantum chemical exchange broadening in methyl groups in proteins. The two relaxation rate constants are established to be linearly dependent using molecular dynamics simulations and empirical analysis of experimental data. This relationship allows chemical exchange broadening to be recognized as an increase in the Hahn-echo relaxation rate constant. The approach is illustrated by analyzing relaxation data collected at three temperatures for E. coli ribonuclease HI and by analyzing relaxation data collected for different cofactor and substrate complexes of E. coli AlkB.
KeywordsAlkB Cross-correlated relaxation Double-quantum relaxation Dynamics Multiple-quantum relaxation Ribonuclease HI Zero-quantum relaxation
Support from National Institutes of Health grants GM089047 (M.L.G.), GM008281 (A. H.). and GM050291 (A.G.P.) is acknowledged gratefully. The AVANCE 600 NMR spectrometer at Columbia University was purchased with the support of NIH grant RR026540. Some of the work presented here was conducted at the Center on Macromolecular Dynamics by NMR Spectroscopy located at the New York Structural Biology Center, supported by a grant from the NIH National Institute of General Medical Sciences (P41 GM118302). A.G.P. is a member of the New York Structural Biology Center. A preliminary account of this work was presented as poster 96 at the 56th Experimental NMR Conference (2016). We thank Richard Friesner (Columbia University) and Martha Beckwith (Advanced Science Research Center, City University of New York) for helpful discussions and Richard Friesner for access to computational facilities. This paper is dedicated to Dennis Torchia (National Institutes of Health) on occasion of his 80th birthday in appreciation of his pioneering achievements in NMR spectroscopy, spin relaxation, and protein dynamics.
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