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The affinity of human RANK binding to its ligand RANKL |
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| Authors: | Shiqian Zhang Changzhen Liu Peng Huang Jingshan Ren Peifu Tang Bin Gao |
| Affiliation: | a The Center for Molecular Immunology, CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China b China-Japan Joint Laboratory of Molecular Immunology and Virology, CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China c Department of Orthopedics, the First Clinical College of Harbin Medical University, Harbin 150001, China d Department of Orthopedics and Trauma Surgery, PLA General Hospital, Beijing 100853, China e Division of Structural Biology and Oxford Protein Production Facility, The Wellcome Trust Centre for Human Genetics, The Henry Wellcome Building for Genomic Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom |
| Abstract: | Receptor activator of nuclear factor-kappa B (RANK) and its ligand, RANKL play critical roles in bone re-modeling, 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 KD value is about 1.09 × 10−10 M. |
| Keywords: | Receptor activator of nuclear factor-kappa B RANK ligand RANK-RANKL interaction Surface plasmon resonance Affinity of RANK-RANKL |
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