title: Structure-activity relationships in human interleukin-1 alpha: identification of key residues for expression of biological activities.
abstract: To identify the sites important for the different biological activities of human interleukin-1 alpha (hIL-1 alpha), 56 single-amino acid-substituted mutants of hIL-1 alpha were produced in Escherichia coli using site-directed mutagenesis, and were examined for their biological activities such as mouse lymphocyte activating factor activity (LAF activity), cytostatic activity against human melanoma cells A-375 (A375 activity) and prostaglandin E2 (PGE2) inducing activity in human osteosarcoma cells MG-63 (PEI activity). Two amino acid residues, Asp26 and Asp151, were found to be important for these activities. The replacement of Asp26 by Val caused a decrease in LAF and PEI activities by one or two orders of magnitude and a slight decrease in A375 activity. The Tyr or Phe substitution for Asp151 caused decreases in LAF and A375 activities by one or two orders of magnitude and complete loss of PEI activity. The change from Asp151 to Lys or Arg resulted in marked decrease in LAF activity and complete loss of A375 and PEI activities. Since Asp26 and Asp151 are close to each other in the three-dimensional structure, the region involving these amino acids seems to be important for the biological activities of hIL-1 alpha.
authors.affiliation: Bioscience Research Laboratories, Dainippon Pharmaceutical Co., Ltd., Osaka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kawashima
authors.forename: J
authors.initials: J
authors.lastname: Yamagishi
authors.forename: M
authors.initials: M
authors.lastname: Yamayoshi
authors.forename: M
authors.initials: M
authors.lastname: Ohue
authors.forename: T
authors.initials: T
authors.lastname: Fukui
authors.forename: H
authors.initials: H
authors.lastname: Kotani
authors.forename: M
authors.initials: M
authors.lastname: Yamada
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003602:Cytotoxicity, Immunologic / Q000235:genetics / Q000502:physiology; D015232:Dinoprostone / Q000096:biosynthesis; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics* / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
chemical_list: D007375:Interleukin-1; D011994:Recombinant Proteins; D015232:Dinoprostone