title: Involvement of the Arg179 in the active site of human IL-6.
abstract: Three internal-amino acid deletions of amino acids 171-179 of human interleukin 6 (IL-6) were introduced at the cDNA level. While all deletion proteins were biologically inactive, immunoprecipitations with a set of conformation-specific anti-(IL-6) monoclonal antibodies showed that only mutant delta 177-179 does not present major alterations in folding. This finding, together with the observation that delta 177-179 is not able to compete with IL-6 for binding to the soluble human IL-6 receptor, suggested that some or all of these three residues participate to the composition of the receptor-binding site of human IL-6. A large number of single-amino-acid-substitution mutants were generated in residues 177, 178 and 179. Their detailed analysis revealed that Arg179 is crucial for activity in mouse cells, because all amino acid substitutions in this position cause a dramatic drop of biological activity on murine hybridoma cells without affecting the overall protein folding. The only substitution which preserved some residual activity was the conservative Arg to Lys change. This demonstrates the absolute requirement for a positive charge in position 179 for the interaction of human IL-6 with its receptor.
authors.affiliation: Institut Pasteur du Brabant, Dept. of Virology, Bruxelles, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Fontaine
authors.forename: R
authors.initials: R
authors.lastname: Savino
authors.forename: R
authors.initials: R
authors.lastname: Arcone
authors.forename: L
authors.initials: L
authors.lastname: de Wit
authors.forename: J P
authors.initials: JP
authors.lastname: Brakenhoff
authors.forename: J
authors.initials: J
authors.lastname: Content
authors.forename: G
authors.initials: G
authors.lastname: Ciliberto
mesh_terms: D001120:Arginine / Q000737:chemistry*; D001483:Base Sequence; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D015870:Gene Expression; D006801:Humans; D007163:Immunosorbent Techniques; D015850:Interleukin-6 / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D015850:Interleukin-6; D011994:Recombinant Proteins; D001120:Arginine