alignment mechanism 通常可以测定的rdc measurements of rdcs.pptVIP

PseudoContact Shifts (PCS) Pseuocontact Shift—PCS Pintacuda etal. Acc Chem Res 2007,Su etal, JACS, 2008/2009 Ln tags: -CYVDTNNDGAYEGDEL PRE Applications Spin label Clore Chem Rev 2009 Tang Nature 2007 Protein conformational exchange Chemical exchange saturation transfer (CEST) 准接触位移 * Long range structural restraints Distance (e- ? H) Distance + Orientation (e- ?H) Mainly Orientation (bond vector) Clore, Chem. Rev. 2009 New Protein NMR Technique Residual Dipolar Couplings Residual Dipolar Couplings (RDC) are measured as contributions to lines splittings in anisotropic solution relative to isotropic solution. RDC核磁实验 残留的偶极耦合的测量是应用以下原理：在不去偶的前提下，先在各向同性条件下（多为水溶液）测定标量耦合常数J，然后在各向异性条件下（即弱定向介质中）测定标量耦合和偶极耦合的总和J+D；偶极耦合D由两种情况的差值得到。 有两类实验：Ecosy类型测定谱峰的裂分；测定一对峰的强度比值 使用不同定向介质的3个理由： 1 同一个耦合常数对应多个角度，所以至少需要2个不同的介质的数据来区分； 2 在某些情况下，尽管有多个核间矢量可以观测到，但是无法得到至少5个独立的成份，这时采用不同的介质可以提供新的约束，虽然新介质导入新的张量，但是这个对于整个刚性分子是公共的，所以还是可能补充新的约束； 3 介质可能会诱导某些构象的取向，但是不太可能若干不同的介质会发生同样的诱导； Alignment is critical to observe RDCs Alignment of a DNA strand with respect to the static magnetic field, B0 Weak alignment can be induced by the intrinsic magnetic susceptibility anisotropy of the molecules Alignment of paramagnetic proteins relative to B0 B0 diagmagnetic paragmagnetic - - - - - - - - - - - - - - - - - - - - Bicelle Bacteriophage Pf1 Alignment of the target molecules is dictated by the interactions between the molecules and the alignment media Better control over the alignment level Multiple aligning conditions External alignment media Purple Membrane Orientation of the uniaxial alignment media is described by a director Alignment mechanism 通常可以测定的RDC Measurements of RDCs Frequency based methods (IPAP) Residual Dipolar Couplings (RDC) are measured as contributions to lines splittings in anisotropic solution relative to isotropic solution. JHNC?的测定:脉冲序列2D IPAP HNCO(Ottiger et al. 1998) JCαC?的测定 ph0=0 ph1=1 ph2=1 3 ph3=3 ph8=0 0 0 0 2 2 2 2 ph30=1 1 1