Thériault Y, Pochapsky T C, Dalvit C, Chiu M L, Sligar S G, Wright P E
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.
J Biomol NMR. 1994 Jul;4(4):491-504. doi: 10.1007/BF00156616.
Sequence-specific backbone 1H and 15N resonance assignments have been made for 95% of the amino acids in sperm whale myoglobin, complexed with carbon monoxide (MbCO). Many assignments for side-chain resonances have also been obtained. Assignments were made by analysis of an extensive series of homonuclear 2D spectra, measured with unlabeled protein, and both 2D and 3D 1H-15N-correlated spectra obtained from uniformly 15N-labeled myoglobin. Patterns of medium-range NOE connectivities indicate the presence of eight helices in positions that are very similar to those found in the crystal structures of sperm whale myoglobin. The resonance assignments of MbCO form the basis for determination of the solution structure and for hydrogen-exchange measurements to probe the stability and folding pathways of myoglobin. They will also form a basis for assignment of the spectra of single-site mutants with altered ligand-binding properties.
已对与一氧化碳结合的抹香鲸肌红蛋白（MbCO）中95%的氨基酸进行了序列特异性主链1H和15N共振归属。还获得了许多侧链共振的归属。归属是通过分析一系列广泛的同核二维谱（用未标记的蛋白质测量）以及从均匀15N标记的肌红蛋白获得的二维和三维1H-15N相关谱来进行的。中程核Overhauser效应（NOE）连接模式表明在与抹香鲸肌红蛋白晶体结构中发现的位置非常相似的位置存在八个螺旋。MbCO的共振归属为确定溶液结构以及进行氢交换测量以探究肌红蛋白的稳定性和折叠途径奠定了基础。它们还将为具有改变的配体结合特性的单点突变体的光谱归属提供基础。
