2D and 3D 15N−13C−13C NMR Chemical Shift Correlation Spectroscopy of Solids: Assignment of MAS Spectra of Peptides Export

@article{citeulike:2828161, abstract = {Strategies are discussed for resolving and assigning peptide backbone and side chain resonances in uniformly 13C,15N-labeled solid peptides. Methods for 2D 13C13C, 15N(13C)13C, and 3D 15N13C13C chemical shift correlation spectroscopy are demonstrated in the chemotactic tripeptide N-formyl-[U-13C,15N]-Met-Leu-Phe-OH (MLF). Band-selective heteronuclear double-cross polarization (DCP) and gamma-encoded homonuclear double-quantum mixing provide large improvements in sensitivity relative to previously published methods. Directional transfers from amide 15N to 13C` or 13Calpha resonances provide two- to 3-fold improvements in signal intensity on the observed 13C spin, in comparison to broadband DCP. Similarly, homonuclear 13C13C transfer is enhanced by use of the rotating frame sequence SPC-5; backbone-to-side chain polarization transfers are achieved with especially high efficiency. Furthermore, the double-quantum nature of the homonuclear transfer permits straightforward classification of C`, Calpha, Cbeta, and Cgamma signals, on the basis of the sign of the cross-peaks. The experiments described here include optimized solid-state analogues of the solution-state schemes commonly employed for amino acid identification and sequential backbone assignment. We expect that these experiments will facilitate application of 2D and 3D correlation methods to assignment of larger solid peptides and proteins.}, address = {Contribution from the Department of Chemistry and Center for Magnetic Resonance, Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139}, author = {Rienstra, Chad M. and Hohwy, Morten and Hong, Mei and Griffin, Robert G.}, citeulike-article-id = {2828161}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ja001092v}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ja001092v}, day = {1}, doi = {10.1021/ja001092v}, journal = {Journal of the American Chemical Society}, keywords = {13c, assignments, mlf, ssnmr}, month = {November}, number = {44}, pages = {10979--10990}, posted-at = {2009-07-08 14:32:20}, priority = {2}, title = {2D and 3D 15N−13C−13C NMR Chemical Shift Correlation Spectroscopy of Solids: Assignment of MAS Spectra of Peptides}, url = {http://dx.doi.org/10.1021/ja001092v}, volume = {122}, year = {2000} }

TY - JOUR ID - citeulike:2828161 L3 - citeulike-article-id:2828161 N2 - Strategies are discussed for resolving and assigning peptide backbone and side chain resonances in uniformly 13C,15N-labeled solid peptides. Methods for 2D 13C13C, 15N(13C)13C, and 3D 15N13C13C chemical shift correlation spectroscopy are demonstrated in the chemotactic tripeptide N-formyl-[U-13C,15N]-Met-Leu-Phe-OH (MLF). Band-selective heteronuclear double-cross polarization (DCP) and gamma-encoded homonuclear double-quantum mixing provide large improvements in sensitivity relative to previously published methods. Directional transfers from amide 15N to 13C` or 13Calpha resonances provide two- to 3-fold improvements in signal intensity on the observed 13C spin, in comparison to broadband DCP. Similarly, homonuclear 13C13C transfer is enhanced by use of the rotating frame sequence SPC-5; backbone-to-side chain polarization transfers are achieved with especially high efficiency. Furthermore, the double-quantum nature of the homonuclear transfer permits straightforward classification of C`, Calpha, Cbeta, and Cgamma signals, on the basis of the sign of the cross-peaks. The experiments described here include optimized solid-state analogues of the solution-state schemes commonly employed for amino acid identification and sequential backbone assignment. We expect that these experiments will facilitate application of 2D and 3D correlation methods to assignment of larger solid peptides and proteins. IS - 44 JF - Journal of the American Chemical Society AD - Contribution from the Department of Chemistry and Center for Magnetic Resonance, Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 EP - 10990 TI - 2D and 3D 15N−13C−13C NMR Chemical Shift Correlation Spectroscopy of Solids: Assignment of MAS Spectra of Peptides VL - 122 SP - 10979 KW - 13c KW - assignments KW - mlf KW - ssnmr AU - Rienstra, Chad AU - Hohwy, Morten AU - Hong, Mei AU - Griffin, Robert PY - 2000/11/01/ UR - http://dx.doi.org/10.1021/ja001092v ER -