Influence of interleukin-6 (IL-6) dimerization on formation of the high affinity hexameric IL-6 receptor complex

被引:48
作者
Ward, LD
Hammacher, A
Howlett, GJ
Matthews, JM
Fabri, L
Moritz, RL
Nice, EC
Weinstock, J
Simpson, RJ
机构
[1] UNIV MELBOURNE,LUDWIG INST CANC RES,WALTER & ELIZA HALL INST MED RES,JOINT PROT STRUCT LAB,PARKVILLE,VIC 3050,AUSTRALIA
[2] UNIV MELBOURNE,DEPT BIOCHEM,PARKVILLE,VIC 3050,AUSTRALIA
[3] LUDWIG INST CANC RES,PARKVILLE,VIC 3050,AUSTRALIA
关键词
D O I
10.1074/jbc.271.33.20138
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 high affinity interleukin-6 (IL-6) signaling complex consists of IL-6 and two membrane-associated receptor components: a low affinity but specific IL-6 receptor and the affinity converter/signal transducing protein gp130. Monomeric (IL-6(M)) and dimeric (IL-6(D)) forms of Escherichia coli-derived human IL-6 and the extracellular (''soluble'') portions of the IL-6 receptor (sIL-6R) and gp130 have been purified in order to investigate the effect of IL-6 dimerization on binding to the receptor complex. Although IL-6(D) has a higher binding affinity for immobilized sIL-6R, as determined by biosensor analysis employing surface plasmon resonance detection, IL-6(M) is more potent than IL-6(D) in a STAT3 phosphorylation assay. The difference in potency is significantly less pronounced when measured in the murine 7TD1 hybridoma growth factor assay and the human hepatoma HepG2 bioassay due to time-dependent dissociation of 37 degrees C of IL-6 dimers into active monomers. The increased binding affinity of IL-6(D) appears to be due to its ability to cross-link two sIL-6R molecules on the biosensor surface. Studies of the IL-6 ternary complex formation demonstrated that the reduced biological potency of IL-6(D) resulted from a decreased ability of the IL-6(D) .(sIL-6R)(2) complex to couple with the soluble portion of gp130. These data imply that the IL-6-induced dimerization of sIL-6R is not the driving force in promoting formation of the hexameric (IL-6 . IL-6R . gp130)(2) complex. A model is presented whereby the trimeric complex of IL-6R, pg130, and IL-6(M) forms before the functional hexamer. Due to its increased affinity for the IL-6R but its decreased ability to couple with gp130, we suggest that a stable IL-6 dimer may be an efficient IL-6 antagonist.
引用
收藏
页码:20138 / 20144
页数:7
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