Homodimerization and heterodimerization of minimal zinc(II)-binding-domain peptides of T-cell proteins CD4, CD8alpha, and Lck.

Homodimerization and heterodimerization of minimal zinc(II)-binding-domain peptides of T-cell proteins CD4, CD8alpha, and Lck.
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T 细胞蛋白 CD4、CD8α 和 Lck 的最小锌 (II) 结合域肽的同二聚化和异二聚化。

DOI:
10.1021/ja9028928
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发表时间:
2009-08-19
影响因子:
15
通讯作者:
Berg, Jeremy M.
Berg, Jeremy M.
中科院分区:
化学1区
文献类型:
--
作者:
Davis, Alisa M.;Berg, Jeremy M.

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金属离子介导的蛋白质寡聚化是一种新兴的蛋白质相互作用模式。T细胞辅助受体CD 4和CD 8 α的C端胞质结构域与激酶Lck的N端区域形成锌桥异二聚体,每个蛋白质为复合物提供两个半胱氨酸配体。使用尺寸排阻色谱法,1H NMR,和UV/可见光吸收光谱与钴(II)作为光谱探针,我们证明,来自这些地区的小肽形成金属桥接的异源二聚体,但也同二聚体,在以前的报告相反。Lck-CD 4和Lck-CD 8 α钴(II)桥接异二聚体复合物比相应的(Lck)2钴(II)复合物分别稳定11 ± 4和22 ± 9倍。这些研究得到以下发现的帮助:钴(II)与钴(II)(-Cys-X-X-Cys-)(-Cys-X-Cys-)发色团的配合物显示出不寻常的光谱,其中可见d至d(4A 2至4 T1(P))跃迁的一个组分发生红移,并且与其他组分很好地分离。这些结果提供了深入了解的基础上的特异性金属桥接的复合物的形成和金属桥接的同源二聚体在T细胞的潜在生物学意义。
Metal-mediated protein oligomerization is an emerging mode of protein-protein interaction. The C-terminal cytosolic domains of T-cell coreceptors CD4 and CD8α form zinc-bridged heterodimers with the N-terminal region of the kinase Lck, with each protein contributing two cysteinate ligands to the complex. Using size exclusion chromatography, 1H NMR, and UV/visible absorption spectroscopy with cobalt(II) as a spectroscopic probe, we demonstrate that small peptides derived from these regions form metal-bridged heterodimers but also homodimers, in contrast to previous reports. The Lck-CD4 and Lck-CD8α cobalt(II)-bridged heterodimer complexes are more stable than the corresponding (Lck)2cobalt(II) complex by factors of 11 ± 4 and 22 ± 9, respectively. These studies were aided by the discovery that cobalt(II) complexes with a cobalt(II)(-Cys-X-X-Cys-)(-Cys-X-Cys-) chromophore show unusual optical spectra with one component of the visible d to d (4A2 to 4T1(P)) transition red-shifted and well separated from the other components. These results provide insights into the basis of specificity of metal-bridged complex formation and on the potential biological significance of metal-bridged homodimers in T-cells.
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