Two-dimensional NMR studies of Kazal proteinase inhibitors. 1. Sequence-specific assignments and secondary structure of turkey ovomucoid third domain.
Two-dimensional NMR studies of Kazal proteinase inhibitors. 1. Sequence-specific assignments and secondary structure of turkey ovomucoid third domain.
复制标题
Kazal 蛋白酶抑制剂的二维 NMR 研究。
DOI：
10.1021/bi00407a039
复制
发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Markley,JL
中科院分区：
文献类型：
--
作者：
Robertson,AD;Westler,WM;Markley,JL
Department of Biochemistry，College of Agricultural and Life Sciences，University of威斯康星州，麦迪逊，威斯康星州53706接收日期：1987年8月11日;修订版：1987年11月24日摘要：二维质子核磁共振实验已被用来顺序分配共振火鸡卵类粘蛋白第三结构域（OMTKY 3）的所有肽骨架质子，除了N-末端的那些。氨基，其信号由于与溶剂交换而未被解析。此外，还对80%以上的侧链质子进行了分析。二维化学位移相关光谱（COSY），中继相干转移光谱（RELAY），和二维homonuclearHartmann-Hahn光谱（HOHAHA）被用来确定自旋系统的几乎一半的残留物之前，顺序分配。两个相邻的残基之间观察到的二维核Overhauser增强的基础上。从另外指定的NOESY交叉峰确定溶液中OMTKY 3的二级结构;其非常类似于通过与灰色链霉菌蛋白酶B复合的OMTKY 3的单晶X射线衍射确定的二级结构[Fujinaga，M.，阅读，R。J.，Sielecki，A.，Ardelt，W.，Laskowski，M.，小的，& James，. N. G. Nat！Acad. Sci. USA 79，4868-4872]。NMR数据提供了三个缓慢交换酰胺质子的证据，这些质子在晶体结构中没有被确定为氢键供体。
Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Madison, Wisconsin 53706 Received August 11, 1987; Revised Manuscript Received November 24, 1987 abstract: Two-dimensional proton NMR experiments havebeen used to sequentially assign resonances to all of the peptide backbone protons of turkey ovomucoid third domain (OMTKY3) except those of the N-terminal-amino group whose signal was not resolved owing to exchange with the solvent. Assignments also have been made for more than 80% of the side-chain protons. Two-dimensional chemical shift correlated spectroscopy (COSY), relayed coherence transfer spectroscopy (RELAY), and two-dimensional homonuclearHartmann-Hahn spectroscopy (HOHAHA) were used to identify the spin systems of almost half of the residues prior to sequential assignment. Assignments were based on two-dimensional nuclear Overhauser enhancements observed between adjacent residues. The secondary structure of OMTKY3 in solution was determined from additional assigned NOESY cross-peaks; it closely resembles the secondary structure determined by single-crystal X-ray diffraction of OMTKY3 in complex with Streptomyces griseus proteinase B [Fujinaga, M., Read, R. J., Sielecki, A., Ardelt, W., Laskowski, M., Jr., & James,. N. G.(1982) Proc. Nat!. Acad. Sci. USA 79, 4868-4872]. The NMR data provide evidence for three slowly exchanging amide protons that were not identified as hydrogen-bond donors in the crystal structure.