5 years ago

Separating Dipolar and Chemical Exchange Magnetization Transfer Processes in 1H-CEST

Separating Dipolar and Chemical Exchange Magnetization Transfer Processes in 1H-CEST
Lewis E. Kay, Tairan Yuwen, Ashok Sekhar
An amide 1H-Chemical Exchange Saturation Transfer (CEST) experiment is presented for studies of conformational exchange in proteins. The approach, exploiting spin-state-selective magnetization transfer, completely suppresses undesired NOE-based dips in CEST profiles so that chemical exchange processes can be studied. The methodology is demonstrated with applications involving proteins that interconvert on the millisecond timescale between major and invisible minor states, and accurate amide 1H chemical shifts of the minor conformer are obtained in each case. The spin-state-selective magnetization transfer approach offers unique possibilities for quantitative studies of protein exchange through 1H-CEST. Finding a needle in a haystack: An amide 1H-CEST experiment for studying conformational exchange processes in proteins is presented. The experiment detects protein millisecond (ms) timescale dynamics through a spin-state-selective scheme that allows the complete suppression of 1H–1H dipolar effects in 1H-CEST profiles that can then be quantified in terms of chemical exchange.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/anie.201610759

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