A PULSED-FIELD GRADIENT ISOTOPE-FILTERED 3D C-13 HMQC-NOESY EXPERIMENT FOR EXTRACTING INTERMOLECULAR NOE CONTACTS IN MOLECULAR-COMPLEXES
A PULSED-FIELD GRADIENT ISOTOPE-FILTERED 3D C-13 HMQC-NOESY EXPERIMENT FOR EXTRACTING INTERMOLECULAR NOE CONTACTS IN MOLECULAR-COMPLEXES
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DOI：
10.1016/0014-5793(94)00740-3
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发表时间：
1994-08-15
期刊：
影响因子：
3.5
通讯作者：
KAY, LE
中科院分区：
文献类型：
--
作者：
LEE, W;REVINGTON, MJ;KAY, LE
用脉冲梯度三维同位素过滤C-13HMQC-NOESY实验研究了37 kDa大分子三元络合物中分子间的相互作用。该络合物由均一的C-13标记色氨酸阻遏因子、天然丰度辅阻遏子、L色氨酸和天然丰度算符DNA组成。该脉冲方案利用脉冲场梯度在同位素过滤过程中去除伪影和消除不需要的磁化，并采用最小化磁化停留在横面上的时间的策略。该实验仅提供了标记蛋白质的质子与未标记物种的质子之间的分子间NOE接触，并已被证明在消除该络合物的同位素编辑的3D C-13 HMQC-NOESY谱中分子内和分子间NOE之间的模糊性方面特别有用。
A pulsed field gradient three-dimensional isotope-filtered C-13 HMQC-NOESY experiment has been developed to characterize intermolecular contacts in a 37 kDa macromolecular ternary complex consisting of uniformly C-13 labeled trp-repressor, its natural abundance co-repressor, L-tryptophan, and natural abundance operator DNA. The pulse scheme makes use of pulsed field gradients for the removal of artifacts and dephasing of unwanted magnetization during isotope filtering, and employs a strategy to minimize the time that magnetization resides in the transverse plane. The experiment provides solely intermolecular NOE contacts between protons of the labeled protein and protons of the unlabeled species, and has proven to be especially useful in eliminating ambiguities between intra- and intermolecular NOEs in the isotope-edited 3D C-13 HMQC-NOESY spectrum of the complex.