Measurement of 15N-1H coupling constants in uniformly 15N-labeled proteins: Application to the photoactive yellow protein |
| |
Authors: | Petra Düx Brian Whitehead Rolf Boelens Robert Kaptein Geerten W Vuister |
| |
Institution: | (1) Department of NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands |
| |
Abstract: | A modified HNHB experiment is presented that allows thedetermination of J(NH) coupling constants directly from the ratio ofcross-peak to diagonal-peak intensities. The experiment was applied to thephotoactive yellow protein (PYP) and yielded the magnitude of 1173J(NH) coupling constants. In addition, 293J(NH(i–1)) coupling constantscould be measured, providing information about the backbone angle .These data, in conjunction with the magnitudes of the3J(HNH) coupling constantsobtained from the HNHA spectrum, effectively discriminate the twopossibilities for the stereospecific assignment of theH resonances in glycine residues. For all eight glycineresidues in PYP that were not subject to conformational averaging and hadnon-degenerate H resonance frequencies, the J-couplingdata, together with limited NOE data, yielded the stereospecific assignmentof the H resonances for these residues. In addition,reliable and precise , dihedral constraints were also derived forthese residues from the J-coupling data. |
| |
Keywords: | J-couplings HNHB HNHA Quantitative J-correlation 3J(NH)" target="_blank">gif" alt="beta" align="MIDDLE" BORDER="0">) 3J(NH(i–" target="_blank">gif" alt="agr" align="BASELINE" BORDER="0">(i– 1)) Stereospecific assignment Glycine residues Photoactive yellow protein |
本文献已被 SpringerLink 等数据库收录! |
|