アブストラクト（33巻2号：The Bulletin of Kanagawa Dental College）

The Bulletin of Kanagawa Dental College

English

Title　： Isolation and Functional Analysis of Baculovirus-expressed Recombinant Human Osteopontin
Subtitle　： ORIGINAL ARTICLES
Authors　： Miwa Horiguchi1), Masahiro Saito2), Masato Yamauchi1), Akira Tsunoda2), Motoshi Suzuki3), Masaharu Takemura3),4), Toshio Teranaka2), Jaro Sodek5), Sadao Sato1)
Authors（kana）　：
Organization　： 1)Department of Craniofacial Growth and Development Dentistry, Kanagawa Dental College, 2)Department of Oral Medicine Division of Operative Dentistry and Endodontics, Kanagawa Dental College, 3)Division of Molecular Carcinogenesis, Center for Neural Disease and Cancer, Nagoya University Graduate School of Medicine, 4)Department of Human Functional Genomics, Functional Genomics Institutes, Life Science Reseach Center, Mie University, 5)CIHR Group in Matrix Dynamics, Faculty of Dentistry, University of Toronto
Journal　： The Bulletin of Kanagawa Dental College
Volume　： 33
Number　： 2
Page　： 61-70
Year/Month　： 2005 / 9
Article　： Original article
Publisher　： Kanagawa Odontological Society
Abstract　： [Abstract] Osteopontin (OPN) is a secreted sialic acid-rich, glycophosphoprotein that is abundant in a non-collagenous matrix of mineralized tissue such as bone. To determine the functional role of OPN in bone formation, a soluble form of recombinant human OPN (rhOPN) was produced through expression in Spodoptera frugiperda (Sf9) insect cells. Purified baculovirus-expressed rhOPN (<2mg/litre of conditioned media) showed biological activities that were more potent than those of the bacterial-expressed rhOPN, as evaluated by the attachment and chemotaxis of osteoblasts. Conversion of the RGD (Arg-Gly-Asp) sequence to KAE (Lys-Ala-Glu) in rhOPN completely eliminated its osteoblast attachment and chemoattractant activity. Immunostaining of vinculin in osteoblasts attached to the baculovirus-expressed rhOPN showed focal contacts consistent with integrin ligation through the RGD sequence. The ability to produce large quantities of biologically-active rhOPN in Sf9 cells will facilitate studies of its biological role in mineralized tissue formation and inflammation and aid in the development of therapeutic applications including for bone tissue engineering.
Practice　： Dentistry
Keywords　： Osteopontin, Cell attachment, Recombinant protein, Osteoblast, Baculovirus