A Methyl‐TROSY‐Based 1H Relaxation Dispersion Experiment for Studies of Conformational Exchange in High Molecular Weight Proteins. Issue 19 (4th April 2019)

• Record Type: Journal Article
• Authors: Yuwen, Tairan; Huang, Rui; Vallurupalli, Pramodh; Kay, Lewis E.
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 19(2019)
• Page Start: 6250
• Page End: 6254
• Publication Date: 2019-04-04
• Subjects: CPMG -- degenerate transitions -- invisible excited states -- methyl NMR -- molecular machines; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201900241
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Molecular complexes often sample conformational states that direct them to specific functions. These states can be difficult to observe through traditional biophysical approaches but they can be studied using a variety of different NMR spin relaxation experiments. However, these applications, when focused on moderate to high molecular weight proteins, are complicated by fast relaxing signals that negatively affect the sensitivity and resolution of spectra. Here a methyl 1 H CPMG‐based experiment for studies of excited conformational states of protein machines is described that exploits a TROSY‐effect to increase signal‐to‐noise. Complexities from the multiplicity of methyl 1 H transitions are addressed to generate a robust pulse scheme that is applied to a 320 kDa homeostasis protein, p97. Abstract : Big wheel keeps on turning : A methyl‐TROSY‐based 1 H SQ‐CPMG experiment for studying μs–ms timescale dynamics in 13 CH3 ‐labeled biomolecules is presented. The experiment preserves slow‐relaxing coherences of 13 CH3 ‐labeled methyl groups, yielding an order of magnitude sensitivity enhancement for studies of high molecular weight proteins relative to other experiments.