title: Trimerization of murine TNF ligand family member LIGHT increases the cytotoxic activity against the FM3A mammary carcinoma cell line.
abstract: LIGHT is a member of the tumor necrosis factor ligand superfamily, which plays important roles in inflammatory and immune responses. LIGHT forms a membrane-anchored homotrimeric complex on the cell surface and is often processed as a soluble protein. Recombinant soluble human LIGHT produced by mammalian cells or Escherichia coli is functional at nanomolar concentrations. However, there is little information about the biological activity of mouse LIGHT (mLIGHT) because of the difficulty in producing bioactive soluble mLIGHT. In this study, recombinant trimeric soluble mLIGHT, or Foldon-mLIGHT, was produced by fusing mLIGHT with the trimerization domain foldon from bacteriophage T4 fibritin. Foldon-mLIGHT was secreted from 293F cells as a 68-kDa trimeric protein. The recombinant protein potently inhibited the growth of the FM3A mouse mammary carcinoma cell line with an IC(50) of 77 pM; however, the monomer or dimer forms of mLIGHT produced by E. coli or mammalian cell systems showed weak or no inhibitory activity. These data clearly indicated that trimerization of soluble mLIGHT is essential for its biological activity.
authors.affiliation: Pharmacology Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 17-85 Jusohonmachi 2-chome, Yodogawa-ku, Osaka, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Ito
authors.forename: Keiji
authors.initials: K
authors.lastname: Iwamoto
authors.forename: Isamu
authors.initials: I
authors.lastname: Tsuji
authors.forename: Hideto
authors.initials: H
authors.lastname: Tsubouchi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Omae
authors.forename: Takayuki
authors.initials: T
authors.lastname: Sato
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Ohba
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Kurokawa
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Taniyama
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shintani
mesh_terms: D000818:Animals; D001681:Biological Assay; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D006801:Humans; D051379:Mice; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000633:toxicity; D012995:Solubility; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000633:toxicity
chemical_list: D011993:Recombinant Fusion Proteins; D053326:Tumor Necrosis Factor Ligand Superfamily Member 14