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Bio-Design and Manufacturing  2021 Vol.4 No.3 P.340~345


Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity

Author(s):  SUN Yi, Marz Pia, Otten Uwe, GE Ji-guang, Rose-John Stefan

Affiliation(s):  College of Life Sciences, Zhejiang University, Hangzhou 310027, China; more

Corresponding email(s):   sun_yi@mail.hz.zj.cn

Key Words:  Ciliary neurotrophic factor(CNTF), Soluble CNTF-Receptor, Fusion protein, Biological activity

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SUN Yi, Marz Pia, Otten Uwe, GE Ji-guang, Rose-John Stefan. Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity[J]. Journal of Zhejiang University Science D, 2021, 4(3): 340~345.

@article{title="Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity",
author="SUN Yi, Marz Pia, Otten Uwe, GE Ji-guang, Rose-John Stefan",
journal="Journal of Zhejiang University Science D",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity
%A Marz Pia
%A Otten Uwe
%A GE Ji-guang
%A Rose-John Stefan
%J Journal of Zhejiang University SCIENCE D
%V 4
%N 3
%P 340~345
%@ 1869-1951
%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2003.0340

T1 - Expression of a fusion protein of human ciliary neurotrophic factor and soluble CNTF-Receptor and identification of its activity
A1 - SUN Yi
A1 - Marz Pia
A1 - Otten Uwe
A1 - GE Ji-guang
A1 - Rose-John Stefan
J0 - Journal of Zhejiang University Science D
VL - 4
IS - 3
SP - 340
EP - 345
%@ 1869-1951
Y1 - 2021
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2003.0340

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 NH2 -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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