PMID: 8960097Nov 29, 1996Paper

Yeast expression of the cytokine receptor domain of the soluble interleukin-6 receptor

Journal of Immunological Methods
P VollmerS Rose-John

Abstract

The complex of the soluble interleukin-6 receptor (sIL-6R) and IL-6 (IL-6) is a potent agonist on cells expressing the signal transducing protein gp 130. In contrast, IL-6 alone only stimulates cells which express a membrane bound form of the IL-6R and gp 130. The natural occurring sIL-6R is generated by shedding of the membrane receptor and to a lesser extend by alternative splicing. We have inserted the coding sequence of the 323 amino acid residues of the human sIL-6R into an expression/secretion vector suitable for the methylotrophic yeast Pichia pastoris. We obtained, however, no detectable expression and secretion of the recombinant protein. When we used only the coding sequence of the cytokine receptor domain of the sIL-6R for the construction of an expression plasmid, this truncated version of the sIL-6R accumulated in the supernatant to 1-5 mg/l. The protein was purified by a single affinity chromatography step using a monoclonal antibody directed against the human IL-6R. Following the same approach, we expressed a truncated splice variant of the sIL-6R. Both, the secreted truncated sIL-6R and the splice variant showed full agonistic biological activity on human hepatoma cells. The described expression strategy will be...Continue Reading

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Citations

Mar 21, 2000·Journal of Neuroscience Research·M ThierS Rose-John
Oct 20, 2005·Journal of Bioscience and Bioengineering·H MizuguchiM Takagi
Oct 20, 2005·Journal of Bioscience and Bioengineering·Kiyoshi YasukawaTakahiko Ishiguro
Feb 1, 1997·Nature Biotechnology·M FischerS Rose-John
Jan 20, 2004·European Journal of Biochemistry·Michael J BeggDeneys R van der Westhuyzen
Mar 28, 1998·The European Journal of Neuroscience·P MärzS Rose-John
Sep 24, 1999·Biological Chemistry·J GrötzingerS Rose-John
Apr 18, 1998·Proceedings of the National Academy of Sciences of the United States of America·P MärzS Rose-John
Apr 9, 1998·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·R K Butlin, T Tregenza
Jan 10, 2001·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·S A JonesG M Fuller

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