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Expressions and purification of a mature form of recombinant human Chemerin in Escherichia coli
Authors:Di Xiang  Jing Zhang  Yizhe Chen  Yiping Guo  Adrian Schalow  Zhonghui Zhang  Xiaojia Hu  Hongjing Yu  Mei Zhao  Shunying Zhu  Huili Lu  Mingyuan Wu  Yan Yu  Anja Moldenhauer  Wei Han
Affiliation:1. Laboratory of Regeneromics, School of Pharmacy, Shanghai Jiaotong University, Shanghai 200240, PR China;2. Institute for Transfusion Medicine, Charite-Universitaetsmedizin Berlin, Campus Virchow-Clinic, Augustenburger Platz 1, 13353 Berlin, Germany;3. Laboratory of Chemistry and Bioactivity of TCM, School of Pharmacy, Shanghai Jiaotong University, Shanghai 200240, PR China;4. Shanghai Municipality Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, PR China;1. Département de Physiologie, and EA 6309 “Maintenance myélinique et neuropathies périphériques”, FR 3503, Facultés de Médecine et de Pharmacie, Université de Limoges, Limoges, F-87025, France;2. Laboratoire de Chimie Analytique et Bromatologie, and EA 3842 “Homéostasie cellulaire et pathologies”, FR 3503, Facultés de Médecine et de Pharmacie, Université de Limoges, Limoges, F-87025, France;1. School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China;2. College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, China;3. Infinitus (China) Company Ltd., Guangzhou 510665, China;4. Center for Translational Medicine, and Shanghai Key Laboratory of Diabetes Mellitus, Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People?s Hospital, Shanghai 200233, China;1. Department of Chemical and Biomolecular Engineering, KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea;2. Institute for the BioCentury, KAIST, Daejeon 305-701, Republic of Korea;1. Department of Chemical Engineering, McGill University, 3610 University Street, Montreal, Quebec H3A 2B2, Canada;2. Department of Mining and Materials Engineering, McGill University, 3610 University Street, Montreal, Quebec H3A 2B2, Canada
Abstract:Chemerin is a novel chemokine that binds to the G protein-coupled receptor (GPCR) ChemR23, also known as chemokine-like receptor 1 (CMKLR1). It is secreted as a precursor and executes pro-inflammatory functions when the last six amino acids are removed from its C-terminus by serine proteases. After maturation, Chemerin attracts dendritic cells and macrophages through binding to ChemR23. We report a new method for expression and purification of mature recombinant human Chemerin (rhChemerin) using a prokaryotic system. After being expressed in bacteria, rhChemerin in inclusion bodies was denatured using 6 M guanidine chloride. Soluble rhChemerin was prepared by the protein-specific renaturation solution under defined conditions. It was subsequently purified using ion-exchange columns to more than 95% purity with endotoxin level <1.0 EU/μg. We further demonstrated its biological activities for attracting migration of human dendritic cells and murine macrophages in vitro using established chemotaxis assays.
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