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Attenuated T 2 relaxation by mutual cancellation of dipole–dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solution

2021; World Scientific; Linguagem: Inglês

10.1142/9789811235795_0006

Konstantin Pervushin, Roland Riek, Gerhard Wider, Kurt Wüthrich,

NMR spectroscopy and applications

NMR with Biological Macromolecules in Solution, pp. 61-66 (2021) No AccessAttenuated T2 relaxation by mutual cancellation of dipole–dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solutionKONSTANTIN PERVUSHIN, ROLAND RIEK, GERHARD WIDER, and KURT WÜTHRICHKONSTANTIN PERVUSHINInstitut für Molekularbiologie und Biophysik Eidgenössische Technische Hochschule Hönggerberg CH-8093 Zurich, Switzerland, ROLAND RIEKInstitut für Molekularbiologie und Biophysik Eidgenössische Technische Hochschule Hönggerberg CH-8093 Zurich, Switzerland, GERHARD WIDERInstitut für Molekularbiologie und Biophysik Eidgenössische Technische Hochschule Hönggerberg CH-8093 Zurich, Switzerland, and KURT WÜTHRICHInstitut für Molekularbiologie und Biophysik Eidgenössische Technische Hochschule Hönggerberg CH-8093 Zurich, Switzerlandhttps://doi.org/10.1142/9789811235795_0006Cited by:0 PreviousNext AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsRecommend to Library ShareShare onFacebookTwitterLinked InRedditEmail Abstract: Fast transverse relaxation of 1H, 15N, and 13C by dipole-dipole coupling (DD) and chemical shift anisotropy (CSA) modulated by rotational molecular motions has a dominant impact on the size limit for biomacromolecular structures that can be studied by NMR spectroscopy in solution. Transverse relaxation-optimized spectroscopy (TROSY) is an approach for suppression of transverse relaxation in multidimensional NMR experiments, which is based on constructive use of interference between DD coupling and CSA. For example, a TROSY-type two-dimensional 1H,15N-correlation experiment with a uniformly 15N-labeled protein in a DNA complex of molecular mass 17 kDa at a 1H frequency of 750 MHz showed that 15N relaxation during 15N chemical shift evolution and 1HN relaxation during signal acquisition both are significantly reduced by mutual compensation of the DD and CSA interactions. The reduction of the linewidths when compared with a conventional two-dimensional 1H,15N-correlation experiment was 60% and 40%, respectively, and the residual linewidths were 5 Hz for 15N and 15 Hz for 1HN at 4°C. Because the ratio of the DD and CSA relaxation rates is nearly independent of the molecular size, a similar percentagewise reduction of the overall transverse relaxation rates is expected for larger proteins. For a 15N-labeled protein of 150 kDa at 750 MHz and 20°C one predicts residual linewidths of 10 Hz for 15N and 45 Hz for 1HN, and for the corresponding uniformly 15N,2H-labeled protein the residual linewidths are predicted to be smaller than 5 Hz and 15 Hz, respectively. The TROSY principle should benefit a variety of multidimensional solution NMR experiments, especially with future use of yet somewhat higher polarizing magnetic fields than are presently available, and thus largely eliminate one of the key factors that limit work with larger molecules. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. © 1997 by The National Academy of Sciences 0027-8424/97/9412366-6$2.00/0 PNAS is available online at http://www.pnas.org. FiguresReferencesRelatedDetails NMR with Biological Macromolecules in SolutionMetrics History PDF download