title: High efficient expression of bioactive human BMP-14 in E. coli using SUMO fusion partner.
abstract: Bone morphogenetic proteins (BMPs) are cytokines from the TGF-β superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, We report here the application of small ubiquitin-related modifier (SUMO) fusion technology to the expression and purification of human BMP-14. The fusion protein expressed in a soluble form was purified to a purity of 90% by Ni-IDA chromatography. After the SUMO-BMP14 fusion protein was cleaved by the SUMO protease at 30 °C for 1 h, the cleaved sample was re-applied to a Ni-IDA. Finally, about 45 mg recombinant hBMP-14 was obtained from 1 litre bacterial culture with no less than 95% purity. The purified hBMP-14 dimer was over 90% purity and could induce the expression of alkaline phosphatase activity in C2C12 cells in a dose-dependent manner. Thus the SUMO-mediated peptide expression and purification system potentially could be employed for the production of other homodimeric proteins.
authors: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing, Jiangsu, People's Republic of China.
authors: Jian Feng
authors: JF
authors: Li
authors: Xian Wei
authors: XW
authors: Cui
authors: Hai Yan
authors: HY
authors: Ji
authors: Ting
authors: T
authors: Qiu
authors: Xue Mei
authors: XM
authors: Ji
authors: Ming Xian
authors: MX
authors: Du
authors: Hai Tao
authors: HT
authors: Wu
authors: Xing Zhou
authors: XZ
authors: Xu
authors: Shuang Quan
authors: SQ
authors: Zhang
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D055428:Growth Differentiation Factor 5 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins