サイトウ マリ   Saito Mari
  齊藤 まり
   所属   鶴見大学  歯学部 歯学科 分子生化学
   職種   助教
論文種別 【査読あり】 研究論文(学術雑誌)
言語種別 英語
査読の有無 査読あり
表題 Protocols for Studying Formation and Mineralization of Dental Tissues In Vivo: Extraction Protocol for Isolating Dentin Matrix Proteins from Developing Teeth.
掲載誌名 正式名:Methods in molecular biology (Clifton, N.J.)
ISSNコード:10643745
出版社 Methods in Molecular Biology
巻・号・頁 1922,pp.239-250
著者・共著者 Yasuo Yamakoshi,Jan C-C Hu,Mari M Saito,James P Simmer
発行年月 2019
概要 The organic material in developing dentin is 90% type I collagen and 10% non-collagenous proteins. The key to understanding dentin biomineralization is to study how these proteins collectively precipitate and organize hydroxyapatite crystals. The first step in characterizing the proteins within a mineralizing matrix is to efficiently extract and isolate the essential molecular participants and elucidate their structural and biochemical properties. In this study, we expanded previous approaches to develop an improved strategy for the extraction of extracellular matrix proteins from the dentin of developing teeth. Proteins in dentin powder were sequentially extracted in the order Tris-guanidine buffer, HCl-formic acid solution, acetic acid-NaCl solution, Tris-NaCl buffer, and a second Tris-guanidine buffer. Individual fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), by gelatin or casein zymography, and by Western blot analysis using dentin sialoprotein (DSP)- or dentin glycoprotein (DGP)-specific antibodies. This approach was used to purify assorted porcine dentin non-collagenous proteins.
DOI 10.1007/978-1-4939-9012-2_24
PMID 30838582
PermalinkURL http://www.scopus.com/inward/record.url?eid=2-s2.0-85062624403&partnerID=MN8TOARS