Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins
Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins
复制标题
DOI：
10.1073/pnas.93.11.5357
复制
发表时间：
1996-05-28
影响因子：
11.1
通讯作者：
Hendrickson, WA
中科院分区：
文献类型：
--
作者：
Lima, CD;Klein, MG;Hendrickson, WA
蛋白激酶C相互作用蛋白1（PKCI-1）的三维结构已经解决了高分辨率的X射线晶体学使用单一的同晶置换与异常散射。通过使用从酵母双杂交系统中鉴定的PKCI-1与蛋白激酶C-β的调节结构域之间的相互作用获得的部分序列，从cDNA文库中克隆编码人PKCI-1的基因。PKCI-1蛋白在毕赤酵母中表达为两个13.7-kDa多肽的二聚体。PKCI-1是HIT蛋白家族的成员，通过序列同一性显示在包括支原体、植物和人类的广泛生物体中是保守的。尽管这种蛋白质序列在自然界中普遍存在，但在体外或体内没有显示出蛋白质产物的独特功能。PKCI-1原聚体具有α + β曲流折叠，其包含五链反平行片层和两个螺旋。两个原聚体聚集在一起形成10链反平行片层，其中一个原聚体的螺旋和羧基末端氨基酸与另一个原聚体中的相应氨基酸之间存在广泛接触。PKCI-1已被证明与锌特异性相互作用。三维结构已经解决了在存在和不存在的锌和两种晶体形式。人PKCI-1的结构提供了该蛋白家族的模型，其表明在整个自然界中保守的稳定折叠。
The three-dimensional structure of protein kinase C interacting protein 1 (PKCI-1) has been solved to high resolution by x-ray crystallography using single isomorphous replacement with anomalous scattering. The gene encoding human PKCI-1 was cloned from a cDNA library by using a partial sequence obtained from interactions identified in the yeast two-hybrid system between PKCI-1 and the regulatory domain of protein kinase C-beta. The PKCI-1 protein was expressed in Pichia pastoris as a dimer of two 13.7-kDa polypeptides. PKCI-1 is a member of the HIT family of proteins, shown by sequence identity to be conserved in a broad range of organisms including mycoplasma, plants, and humans. Despite the ubiquity of this protein sequence in nature, no distinct function has been shown for the protein product in vitro or in vivo. The PKCI-1 protomer has an alpha+beta meander fold containing a five-stranded antiparallel sheet and two helices. Two protomers come together to form a 10-stranded antiparallel sheet with extensive contacts between a helix and carboxy terminal amino acids of a protomer with the corresponding amino acids in the other protomer. PKCI-1 has been shown to interact specifically with zinc. The three-dimensional structure has been solved in the presence and absence of zinc and in two crystal forms. The structure of human PKCI-1 provides a model of this family of proteins which suggests a stable fold conserved throughout nature.