title: Domain 5 of high molecular weight kininogen inhibits collagen-mediated cancer cell adhesion and invasion in association with α-actinin-4.
abstract: High molecular weight kininogen (HK) is a plasma glycoprotein with multiple functions, including the regulation of coagulation. We previously demonstrated that domain 5 (D5(H)), a functional domain of HK, and its derived peptides played an important role in the vitronectin-mediated suppression of cancer cell adhesion and invasion. However, the underlying mechanisms of the D5(H)-mediated suppressive effects remain to be elucidated. Here, we showed that D5(H) and its derivatives inhibited the collagen-mediated cell adhesion and invasion of human osteosarcoma MG63 cells. Using purified D5(H) fused to glutathione-S-transferase (GST) and D5(H)-derived peptides for column chromatography, an actin-binding protein, α-actinin-4, was identified as a binding protein of D5(H) with high-affinity for P-5m, a core octapeptide of D5(H). Immunofluorescence microscopy demonstrated that D5(H) co-localized with α-actinin-4 inside MG63 cells. In addition, exogenous GST-D5(H) added to the culture media was transported into MG63 cells, although GST alone as a control was not. As α-actinin-4 regulates actin polymerization necessary for cell adhesion and is related to the integrin-dependent attachment of cells to the extracellular matrix, our results suggest that D5(H) may modulate cell adhesion and invasion together with actinin-4.
authors.affiliation: Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan.
authors.forename: Tsunetoshi
authors.initials: T
authors.lastname: Hatoh
authors.forename: Toshinaga
authors.initials: T
authors.lastname: Maeda
authors.forename: Keisuke
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Osamu
authors.initials: O
authors.lastname: Ogikubo
authors.forename: Susumu
authors.initials: S
authors.lastname: Uchiyama
authors.forename: Takanobu
authors.initials: T
authors.lastname: Otsuka
authors.forename: Iwao
authors.initials: I
authors.lastname: Ohkubo
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Ogita
mesh_terms: D000185:Actinin / Q000378:metabolism*; D000595:Amino Acid Sequence; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D003094:Collagen / Q000378:metabolism; D006801:Humans; D019679:Kininogen, High-Molecular-Weight / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: C111453:ACTN4 protein, human; D019679:Kininogen, High-Molecular-Weight; D010446:Peptide Fragments; D000185:Actinin; D003094:Collagen