Molecular modeling-guided mutagenesis of the extracellular part of gp130 leads to the identification of contact sites in the interleukin-6 (IL-6)center dot IL-6 receptor center dot gp130 complex

被引:63
作者
Horsten, U
MullerNewen, G
Gerhartz, C
Wollmer, A
Wijdenes, J
Heinrich, PC
Grotzinger, J
机构
[1] RHEIN WESTFAL TH AACHEN,INST BIOCHEM,D-52057 AACHEN,GERMANY
[2] DIACLONE,F-25020 BESANCON,FRANCE
关键词
D O I
10.1074/jbc.272.38.23748
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The transmembrane protein gp130 is involved in many cytokine-mediated cellular responses and acts therein as the signal-transducing subunit, In the case of interleukin-6 (IL-6), the signal-transducing complex is composed of the ligand IL-6, the IL-6 receptor (IL-6R, gp80, CD126), and at least two gp130 (CD130) molecules, The extracellular part of the signal transducer gp130 consists of sis fibronectin type III-like domains, It has recently been shown that the three membrane distal domains bind to the IL-6.IL-6R complex. A structural model of the IL-6.IL-6R gp130 complex enabled us to propose amino acid residues in these domains of gp130 interacting with IL-6 bound to its receptor. The proposed amino acid residues located in the B'C' loop (Val(252)) and in the F'G' loop (Gly(306), Lys(307)) of domain 3 and in the hinge region (Tyr(21S)) connecting domains 2 and 3 of gp130 were mutated to disturb ternary complex formation, Binding of wild type and mutants of the extracellular region of gp130 was studied by use of a co-precipitation assay and Scatchard analysis, All mutants showed decreased binding to the IL-6.IL-6R complex, Biological function of the membrane bound gp130 mutants was studied by STAT (signal transducer and activator of transcription) activation in COS-7 cells and by proliferation of stably transfected Ba/F3 cells. Reduced binding of the mutants was accompanied by decreased biological activity, The combined approach of molecular modeling and site-directed mutagenesis has led to the identification of amino acid residues in gp130 required for complex formation with IL-6 and its receptor.
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页码:23748 / 23757
页数:10
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