Simultaneous measurement of protein one-bond and two-bond nitrogen-carbon coupling constants using an internally referenced quantitative J-correlated [p>15p>N,p>1p>H]-TROSY-HNC

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• 作者：Wienk ; Hans L.J. ; Martí ; nez ; Mitcheell M. ; Yalloway ; Gary N. ; Schmidt ; Jü ; rgen M. ; P&eacute ; rez ; Carlos ; Rü ; terjans ; Heinz ; Lö ; hr ; Frank
• 关键词：DFPase ; flavodoxin ; perdeuteration ; residual dipolar coupling ; scalar coupling ; TROSY ; xylanase
• 刊名：Journal of Biomolecular NMR
• 出版年：2003
• 出版时间：2003
• 年：2003
• 卷：25
• 期：2
• 页码：133-145
• 全文大小：197 KB

文摘

A quantitative J-correlation pulse sequence is described that allows simultaneous determination of one-bond and two-bond nitrogen-carbon coupling constants for protonated or deuterated proteins. Coupling constants are calculated from volume ratios between cross peaks and reference axial peaks observed in a single 3D spectrum. Accurate backbone p>1p>J_NC′, p>1p>J_NC&agr;, and p>2p>J_NC&agr; coupling constants are obtained for the two [p>15p>N;p>13p>C]-labeled, medium-sized proteins flavodoxin and xylanase and for the [p>2p>H;p>15p>N;p>13p>C]-labeled, large protein DFPase. A dependence of one-bond and two-bond J_NC&agr; values on protein backbone &psgr; torsion angles is readily apparent, in agreement with previously found correlations. In addition, the experiment is performed on isotropic as well as aligned protein to measure associated p>15p>N-p>13p>C residual dipolar couplings.p>