| Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity |
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| 作者姓名： | 孙益 Mrz Pia Otten Uwe 葛霁光 Rose-John Stefan |
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| 作者单位： | College of Life Sciences，Zhejiang University，Department of Physiology，Basel University，Department of Physiology，Basel University，College of Life Sciences，Zhejiang University，Department of Biochemistry，Christian Albrechts University of Kiel，D Hangzhou 310027，China，Vesalgasse 1，4051 Basel，Switzerland，Vesalgasse 1，4051 Basel，Switzerland，Hangzhou 310027，China，24098 Kiel，Germany |
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| 基金项目： | ProjectsupportedbytheScientificResearchFoundationfortheReturnedOverseasChineseScholar，ZhejiangProvince，ChinaandSFB415，TPB5，Germany . |
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| 摘 要： | INTRODUCTIONCiliaryneurotrophicfactor (CNTF)isanat urallyoccurringproteinwithamolecularmassofapproximately 2 2kDa .Itisexpressedinglialcellswithinthecentralandperipheralnervoussystems.CNTFstimulatesgeneexpression ,cellsurvivalordifferentiationinavarietyof…
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Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity |
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| Abstract: | 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 gp130 and LIF R, forming a tripartite receptor complex. Cells that only express gp130 and LIF R, but not CNTF R are refractory to stimulation by CNTF. On many target cells 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 NH 2 terminus of CNTF. Recombinant CNTF/sCNTF R fusion protein (Hyper CNTF) was successfully 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 transfected BAF/3 cells in response to CNTF and Hyper CNTF were used to verify the activity of the cytokines. The proliferative results confirmed that CNTF required homodimerization of the gp130, CNTF R and LIF R receptor subunit whereas Hyper CNTF required heterodimerization of the gp130 and LIF R receptor subunit. We concluded that the fusion protein Hyper CNTF had superagonistic activity on target cells expressing gp130 and LIF R, but lacking membrane bound CNTF R. |
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| Keywords: | Ciliary neurotrophic factor(CNTF) Soluble CNTF Receptor Fusion protein Biological activity |
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