title: Optimization of the inter-domain structure of galectin-9 for recombinant production.
abstract: We previously developed a stable form of galectin-9, an immunomodulatory animal lectin with a truncated linker peptide (G9Null), to overcome the protease sensitivity of wild-type galectin-9. G9Null is highly resistant to proteolysis, while the modification marginally improved the low solubility of the wild-type protein. To increase its solubility, we further modified the remaining linker region of G9Null. A 10-amino acid deletion with a single amino acid substitution resulted in an ∼400% increase in solubility and yield without an adverse effect on its biological activity. This mutant protein might be useful for large-scale recombinant production needed for evaluation of the therapeutic potential of galectin-9.
authors.affiliation: Division of Research Instrument and Equipment, Life Science Research Center, Japan.
authors.forename: Aiko
authors.initials: A
authors.lastname: Itoh
authors.forename: Yoko
authors.initials: Y
authors.lastname: Fukata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Miyanaka
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Nonaka
authors.forename: Takashi
authors.initials: T
authors.lastname: Ogawa
authors.forename: Takanori
authors.initials: T
authors.lastname: Nakamura
authors.forename: Nozomu
authors.initials: N
authors.lastname: Nishi
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D037161:Galectins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012995:Solubility
chemical_list: D037161:Galectins; D011993:Recombinant Fusion Proteins
keywords: Galectin; Linker peptide; Protein engineering; Therapeutic agent