Solution structures in SDS micelles and functional activity at the bullfrog substance P receptor of ranatachykinin peptides.
Solution structures in SDS micelles and functional activity at the bullfrog substance P receptor of ranatachykinin peptides.
复制标题
DOI：
10.1021/jm000093v
复制
发表时间：
2000-04
影响因子：
7.3
通讯作者：
S. Perrine;T. Whitehead;R. Hicks;J. Szarek;J. Krause;M. A. Simmons
中科院分区：
文献类型：
--
作者：
S. Perrine;T. Whitehead;R. Hicks;J. Szarek;J. Krause;M. A. Simmons
从牛蛙脑和肠道中分离到一组新的速激肽样肽。这些化合物，即Ranatachykinin A（RTKA）、Ranatachykinin B（RTKB）和Ranatachykinin C（RTKC），以其来源牛蛙（Rana catesbeiana）及其与速激肽肽家族的同源性命名。我们提出的胶束结合的结构和药理作用的RTK的第一份报告。使用（1）H NMR化学位移分配、二维NMR技术、分子动力学和模拟退火程序在膜模型环境中生成RTK的三维结构，从而可以确定可能的预结合配体构象。RTKA、RTKB和RTKC被确定为从中间区域到C末端（残基4-10）是螺旋状的，在N末端具有较大程度的柔性，在C末端具有较小的动态磨损。RTKs的药理学作用通过测量稳定转染牛蛙P物质受体（bfSPR）的中国仓鼠卵巢细胞中细胞内Ca（2+）的升高来研究。所有RTKs均引起Ca（2+）升高，最大效应的顺序为RTKA >/= SP > RTKC >/= RTKB。高浓度（1 μ M）的神经肽对随后使用相同或不同肽的激发产生不同程度的脱敏，而低浓度（1 μ M）在bfSPR处产生致敏作用。我们的数据表明，氨基酸侧链的差异和它们在C-末端序列的电荷状态或在N-末端的二级结构的差异，根据本文的研究结果不重叠，可以解释不同程度和类型的受体激活在bfSPR。
A set of novel tachykinin-like peptides has been isolated from bullfrog brain and gut. These compounds, ranatachykinin A (RTKA), ranatachykinin B (RTKB), and ranatachykinin C (RTKC), were named for their source, Rana catesbeiana, and their homology to the tachykinin peptide family. We present the first report of the micelle-bound structures and pharmacological actions of the RTKs. Generation of three-dimensional structures of the RTKs in a membrane-model environment using (1)H NMR chemical shift assignments, two-dimensional NMR techniques, and molecular dynamics and simulated annealing procedures allowed for the determination of possible prebinding ligand conformations. RTKA, RTKB, and RTKC were determined to be helical from the midregion to the C-terminus (residues 4-10), with a large degree of flexibility in the N-terminus and minor dynamic fraying at the end of the C-terminus. The pharmacological effects of the RTKs were studied by measuring the elevation of intracellular Ca(2+) in Chinese hamster ovarian cells stably transfected with the bullfrog substance P receptor (bfSPR). All of the RTKs tested elicited Ca(2+) elevations with a rank order of maximal effect of RTKA >/= SP > RTKC >/= RTKB. A high concentration (1 microM) of the neuropeptides produced varying degrees of desensitization to a subsequent challenge with the same or different peptide, while a low concentration (1 pM) produced sensitization at the bfSPR. Our data suggest differences in amino acid side chains and their charged states at the C-terminal sequence or differences in secondary structure at the N-terminus, which do not overlap according to the findings in this paper, may explain the differing degree and type of receptor activation seen at the bfSPR.