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. 1997 Aug 1;325(Pt 3):693–700. doi: 10.1042/bj3250693

Evidence for the formation of a heterotrimeric complex of leukaemia inhibitory factor with its receptor subunits in solution.

J G Zhang 1, C M Owczarek 1, L D Ward 1, G J Howlett 1, L J Fabri 1, B A Roberts 1, N A Nicola 1
PMCID: PMC1218613  PMID: 9271090

Abstract

Leukaemia inhibitory factor (LIF) is a polyfunctional cytokine that is known to require at least two distinct receptor components (LIF receptor alpha-chain and gp130) in order to form a high-affinity, functional, receptor complex. Human LIF binds with unusually high affinity to a naturally occurring mouse soluble LIF receptor alpha-chain, and this property was used to purify a stable complex of human LIF and mouse LIF receptor alpha-chain from pregnant-mouse serum. Recombinant soluble human gp130 was expressed, with a FLAG(R) epitope (DYKDDDDK) at the N-terminus, in the methylotropic yeast Pichia pastoris and purified using affinity chromatography. The formation of a trimeric complex in solution was established by native gel electrophoresis, gel-filtration chromatography, sedimentation equilibrium analysis, surface plasmon resonance spectroscopy and chemical cross-linking. The stoichiometry of this solution complex was 1:1:1, in contrast with that of the complex of interleukin-6, the interleukin-6-specific low-affinity receptor subunit and gp130, which is 2:2:2.

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Selected References

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