| Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity |
| 陈益[1] MārzPia[2] OttenUwe[2] 葛霁光[1] Rose-JohnStefan[3] |
| 关键词：融合蛋白 基因表达 可溶性CNTF-受体 活性鉴别 人体纤毛状亲神经因子 CNTF |
| 主要内容：Ciliary neurotrophic factor (CNTF) has pleiotropic actions on many neuronal populations as well as on glia. Signal transduction by CNTF requires that it bind first to CNTF-R, permitting the recruitment of gpl30 and LIF-R, forming a tripartite receptor complex. Ceils that only express gpl30 and LIF-R, but not CNTF-R are refractory to stimulation by CNTF. On many target ceils CNTF only acts in the presence of its specific agonistic soluble receptors. We engineered a soluble fusion protein by linking the COOH-terminus of sCNTF-R to the NH2-terminus of CNTF. Recombinant CNTF/sCNTF-R fusion protein (Hyper-CNTF) was sac-cessfully expressed in COS-7 cells. The apparent molecular mass of the Hyper-CNTF protein was estimated from western blots to be 75 kDa. Proliferation assays of tmnsfected BAF/3 cells in response to CNTF and Hy-per-CNTF were used to verify the activity of the cytokines. The proliferative results confirmed that CNTF required homodimerization of the gpl30, CNTF-R and LIF-R receptor subunlt whereas Hyper-CNTF required heterodimerization of the gpl30 and LIF-R receptor subunit. We concluded that the fusion protein Hyper-CNTF had superagonistic activity on target cells expressing gpl30 and LIF-R, but lacking membrane-beund CNTF-R. |
| 《浙江大学学报：自然科学英文版》 2003,4(3).-340-345 |
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