title: An isotope labeling strategy for methyl TROSY spectroscopy.
abstract: Recently we have shown that HMQC spectra of protonated methyl groups in high molecular weight, highly deuterated proteins have large enhancements in sensitivity and resolution relative to HSQC-generated data sets. These enhancements derive from a TROSY effect in which complete cancellation of intra-methyl (1)H-(1)H and (1)H-(13)C dipolar interactions occurs for 50% of the signal in the case of HMQC, so long as the methyl is attached to a molecule tumbling in the macromolecular limit (Tugarinov, V., Hwang, P.M., Ollerenshaw, J.E., Kay, L.E. J. Am. Chem. Soc. (2003) 125, 10420-10428; Ollerenshaw, J.E., Tugarinov, V. and Kay, L.E. Magn. Reson. Chem. (2003) 41, 843-852. The first demonstration of this effect was made for isoleucine delta1 methyl groups in a highly deuterated 82 kDa protein, malate synthase G. As with (1)H-(15)N TROSY spectroscopy high levels of deuteration are critical for maximizing the TROSY effect. Here we show that excellent quality methyl TROSY spectra can be recorded on U-[(2)H] Iledelta1-[(13)CH(3)] Leu,Val-[(13)CH(3)/(12)CD(3)] protein samples, significantly extending the number of probes available for structural and dynamic studies of high molecular weight systems.
authors.affiliation: Protein Engineering Network Centres of Excellence and the Department of Medical Genetics, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
authors.forename: Vitali
authors.initials: V
authors.lastname: Tugarinov
authors.forename: Lewis E
authors.initials: LE
authors.lastname: Kay
mesh_terms: D002247:Carbon Isotopes; D003903:Deuterium; D008745:Methylation; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D003903:Deuterium