人网织钙结合蛋白-1中依赖钙的结构变化
Calcium-dependent structural changes in human reticulocalbin-1.
作者信息
Suzuki Nanao, Ban Syoko, Itoh Eriko, Chen Sisi, Imai Fabiana L, Sawano Yoriko, Miyakawa Takuya, Tanokura Masaru, Yonezawa Naoto
机构信息
Department of Chemistry, Graduate School of Science, Chiba University, Chiba 263-8522, Japan; Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan; and Laboratory of Chemistry, College of Liberal and Sciences, Tokyo Medical and Dental University, Chiba 272-0827, Japan.
出版信息
J Biochem. 2014 May;155(5):281-93. doi: 10.1093/jb/mvu003. Epub 2014 Jan 21.
Human reticulocalbin-1 (hRCN1) has six EF-hand motifs and binds Ca(2+). hRCN1 is a member of the CREC family localized in the secretory pathway, and its cellular function remains unclear. In this study, we established a new bacterial expression and purification procedure for hRCN1. We observed that hRCN1 binds Ca(2+) in a cooperative manner and the Ca(2+) binding caused an increase in the α-helix content of hRCN1. On the other hand, hRCN1 did not change the structure with Mg(2+) loading. hRCN1 is a monomeric protein, and its overall structure became more compact upon Ca(2+) binding, as revealed by gel-filtration column chromatography and small-angle X-ray scattering. This is the first report of conformational changes in the CREC family upon Ca(2+) binding. Our data suggest that CREC family member interactions with target proteins are regulated in the secretory pathway by conformational changes upon Ca(2+) binding.
人网织钙结合蛋白-1（hRCN1）有六个EF手基序并能结合Ca²⁺。hRCN1是定位于分泌途径的CREC家族成员，其细胞功能尚不清楚。在本研究中，我们建立了一种新的hRCN1细菌表达和纯化方法。我们观察到hRCN1以协同方式结合Ca²⁺，且Ca²⁺结合导致hRCN1的α-螺旋含量增加。另一方面，hRCN1在加载Mg²⁺时结构不变。hRCN1是一种单体蛋白，凝胶过滤柱色谱和小角X射线散射显示，其整体结构在结合Ca²⁺后变得更加紧凑。这是关于CREC家族在结合Ca²⁺后构象变化的首次报道。我们的数据表明，CREC家族成员与靶蛋白的相互作用在分泌途径中通过结合Ca²⁺后的构象变化来调节。
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