The affinity of human RANK binding to its ligand RANKL
The affinity of human RANK binding to its ligand RANKL
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人 RANK 与其配体 RANKL 结合的亲和力
DOI：
10.1016/j.abb.2009.04.008
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发表时间：
2009-07-01
影响因子：
3.9
通讯作者：
Gao, Bin
中科院分区：
文献类型：
--
作者：
Zhang, Shiqian;Liu, Changzhen;Gao, Bin
核因子-kappaB受体激活剂RANK及其配体RANKL在骨重建、免疫功能、血管疾病和乳腺发育中起重要作用。为了研究RANK和RANKL的相互作用，我们利用大肠杆菌表达系统表达了RANKL的胞外区和胞外区。RANK首先以包涵体形式表达，随后适当复性，而RANKL最初以GST融合蛋白的形式表达，然后通过酶切去除GST。凝胶过滤层析和交联实验表明，RANKL在溶液中以单体形式存在，而RANKL则以三聚体形式存在。用表面等离子体共振技术测定RANKL与RANKL的结合亲和力，K-D值约为1.09×10~(-10)，M.(C)2009 Elsevier Inc.
Receptor activator of nuclear factor-kappa B (RANK) and its ligand, RANKL play critical roles in bone remodeling, immune function, vascular disease and mammary gland development. To Study the interaction of RANK and RANKL, we have expressed both extracellular domain of RANK and ectodomain of RANKL using Escherichia coli expression system. RANK was expressed as an inclusion body first which Properly refolded later, while RANKL was initially produced as a GST fusion protein, after which the GST was removed by enzyme digestion. Soluble RANK existed as a monomer while RANKL was seen as a trimer in solution, demonstrated by gel filtration chromatography and cross-linking experiment. The recombinant RANK and RANKL Could bind to each other and the binding affinity of RANKL for RANK was measured with Surface plasmon resonance technology and K-D value is about 1.09 x 10(-10), M. (C) 2009 Elsevier Inc. All rights reserved.