CHARACTERIZATION OF THE 3-DIMENSIONAL SOLUTION STRUCTURE OF HUMAN PROFILIN - H-1, C-13, AND N-15 NMR ASSIGNMENTS AND GLOBAL FOLDING PATTERN
CHARACTERIZATION OF THE 3-DIMENSIONAL SOLUTION STRUCTURE OF HUMAN PROFILIN - H-1, C-13, AND N-15 NMR ASSIGNMENTS AND GLOBAL FOLDING PATTERN
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DOI：
10.1021/bi00213a010
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发表时间：
1993-12-21
期刊：
影响因子：
2.9
通讯作者：
MUELLER, L
中科院分区：
文献类型：
--
作者：
METZLER, WJ;CONSTANTINE, KL;MUELLER, L
人profilin是一种15 kDa的蛋白质，在导致细胞骨架重排的信号通路中起主要作用。基本上完整的分配的H-1，C-13，和N-15共振的人profilin已通过分析多维，双和三重共振核磁共振（NMR）实验。分析了C-13 α和C-13 β化学位移与其各自的无规卷曲值的偏差，并与从NMR数据确定的二级结构良好相关。在X-PLOR程序中使用总共1186个实验衍生的构象约束对20个人profilin结构进行了优化。对于骨架原子，结构收敛到1.5埃的均方根距离偏差。所得构象系综表明，人profilin是一种α/β蛋白，由七链、反向平行的β折叠和三个螺旋组成。人profilin的二级结构元件与在阿米巴profilin I中发现的那些非常相似[Archer，S. J.，文森，V.K.，Pollard，T. D、& Torchia，D. A.（1993），Biochemistry 32，6680-6687]，这表明阿米巴profilin I的三维结构应该类似于此处确定的人profilin的三维结构。人profilin的结构测定促进了低等真核生物和人profilin的序列比对，并提供了一个框架，在此基础上可以探索profilin的各种功能。人促凝血酶原蛋白肌动蛋白结合区的至少一个元件是α-螺旋。磷脂酰肌醇4，5-二磷酸可以干扰肌动蛋白结合人profilin提出了两种机制。
Human profilin is a 15-kDa protein that plays a major role in the signaling pathway leading to cytoskeletal rearrangement. Essentially complete assignment of the H-1, C-13, and N-15 resonances of human profilin have been made by analysis of multidimensional, double- and triple-resonance nuclear magnetic resonance (NMR) experiments. The deviation of the C-13alpha and C-13beta chemical shifts from their respective random coil values were analyzed and correlate well with the secondary structure determined from the NMR data. Twenty structures of human profilin were refined in the program X-PLOR using a total of 1186 experimentally derived conformational restraints. The structures converged to a root mean squared distance deviation of 1.5 angstrom for the backbone atoms. The resultant conformational ensemble indicates that human profilin is an alpha/beta protein comprised of a seven-stranded, antiparallel beta-sheet and three helices. The secondary structure elements for human profilin are quite similar to those found in Acanthamoeba profilin I [Archer, S. J., Vinson, V. K., Pollard, T. D., & Torchia, D. A. (1993), Biochemistry 32, 6680-6687], suggesting that the three-dimensional structure of Acanthamoeba profilin I should be analogous to that determined here for human profilin. The structure determination of human profilin has facilitated the sequence alignment of lower eukaryotic and human profilins and provides a framework upon which the various functionalities of profilin can be explored. At least one element of the actin-binding region of human profilin is an alpha-helix. Two mechanisms by which phosphatidylinositol 4,5-bisphosphate can interfere with actin-binding by human profilin are proposed.