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首页> 外文期刊>Biochemistry >Metal-thiolate clusters in the C-terminal domain of human neuronal growth inhibitory factor (GIF)
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Metal-thiolate clusters in the C-terminal domain of human neuronal growth inhibitory factor (GIF)

机译：人神经元生长抑制因子（GIF）C端结构域中的金属硫醇盐簇

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摘要

Neuronal growth inhibitory factor (GIF), a metallothionein-like protein (metallothionein-3), impairs the survival and neurite formation of cultured neurons. Native GIF contains 4 Cu(I) and three Zn(ZI) ions organized in homometallic metal-thiolate clusters. However, the cluster localization is not known. In this study, the metal-thiolate clusters formed with monovalent and divalent metal ions in the C-terminal domain of human GIF [GIF(32-68)] containing 11 cysteines were investigated. The cluster formation was followed by using electronic absorption, circular dichroism (CD), and magnetic circular dichroism (MCD) spectroscopy, and in the case of Cu(I) complexes also by luminescence spectroscopy at 77 K. Spectroscopic studies on the Cu(I)-GIF(32-68) complexes showed the successive formation of two air-sensitive Cu4S8-9- and Cu6S11-clusters. With Zn(II) and Cd(II) ions, a well-defined M4S11-cluster is formed in which each metal ion is tetrahedrally coordinated by cysteine thiolates. In the Cd-113 NMR spectra of Cd-113(4)-GIF(32-68), recorded at 293 and 323 K, all four Cd-113 resonances at 672.8, 620.9, 629.6, and 564.2 ppm were observed only at 323 K. Their detection at elevated temperature indicates a conformational flexibility of this domain. Evidence for the existence of a Cds-GIF(32-68) complex, contaning two more weakly bound Cd(II) ions, was also obtained. The formation of this complex requires the transformation of some originally terminal thiolates of the Cd4S11-cluster to bridging thiolates, suggesting a more accessible cluster structure. Such properties of Cd-4-GIF(32-68) have not been observed with the Cd4S11-cluster in the isolated cr-domain (amino acids 31-61) of metallothioneins. The significance of Cu- and Zn-clusters for the structure of native GIF is discussed. [References: 48]
机译：神经元生长抑制因子（GIF）是一种金属硫蛋白样蛋白（metallothionein-3），会损害培养的神经元的存活和神经突的形成。天然GIF包含4个Cu（I）和3个Zn（ZI）离子，它们组织在均金属硫醇盐簇中。但是，群集本地化是未知的。在这项研究中，研究了在含有11个半胱氨酸的人GIF [GIF（32-68）] C端结构域中与一价和二价金属离子形成的金属硫醇盐簇。在形成团簇之后，使用电子吸收，圆二色性（CD）和磁圆二色性（MCD）光谱，对于Cu（I）配合物，还通过77 K的发光光谱进行研究。Cu（I）的光谱研究）-GIF（32-68）配合物显示了两个对空气敏感的Cu4S8-9-和Cu6S11簇的连续形成。利用Zn（II）和Cd（II）离子，形成了定义明确的M4S11簇，其中每个金属离子都被半胱氨酸硫醇盐四面体配位。在293和323 K记录的Cd-113（4）-GIF（32-68）的Cd-113 NMR光谱中，仅在323处观察到672.8、620.9、629.6和564.2 ppm的所有四个Cd-113共振K.在高温下检测到它们表明该结构域的构象柔性。还获得了包含两个较弱结合的Cd（II）离子的Cds-GIF（32-68）配合物存在的证据。此复合物的形成需要将Cd4S11-簇的一些最初末端的硫醇盐转变为桥连的硫醇盐，这表明簇结构更易获得。在金属硫蛋白的分离的cr-结构域（氨基酸31-61）中，没有用Cd4S11-簇观察到Cd-4-GIF（32-68）的这种性质。讨论了Cu和Zn簇对于天然GIF结构的重要性。 [参考：48]

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