title: Expression and purification of soluble human APRIL in Escherichia coli using ELP-SUMO tag.
abstract: APRIL is a member of the tumor necrosis factor (TNF) family of ligands that mediate tumor cells proliferation as well as survival, depending on the cellular context. In this report, we present a novel method to obtain soluble human APRIL in Escherichia coli using the elastin-like polypeptide and SUMO (ELP-SUMO) tags. The fusion protein with ELP-SUMO tag was expressed in a soluble form at 15°C. After purification based on inverse transition cycling (ITC) method, the purified ELP-SUMO-hAPRIL fusion protein was subsequently cleaved by SUMO protease to release mature hAPRIL. Following affinity chromatography, the target protein was re-purified with high purity. Finally, about 4.8mg recombinant hAPRIL was obtained from 1l bacterial culture with no less than 85% purity. The molecular mass (Mr) of the recombinant hAPRIL was confirmed by MALDI-TOF MS as Mr 16,314. The purified hAPRIL exhibits biological activity on Jurkat cells. It is the first report on soluble production of hAPRIL in E. coli using ELP-SUMO tag.
authors.affiliation: Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou, Jiangsu Province, PR China; Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing 210046, Jiangsu, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing 210046, Jiangsu, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing 210046, Jiangsu, PR China. Electronic address: shuangquanz@yahoo.com.
authors.forename: Shuang Quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D003001:Cloning, Molecular; D004549:Elastin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D025842:SUMO-1 Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D053300:Tumor Necrosis Factor Ligand Superfamily Member 13 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; D053300:Tumor Necrosis Factor Ligand Superfamily Member 13; D004549:Elastin
keywords: ELP-SUMO; Escherichia coli; Soluble expression; hAPRIL