title: Characterization of mineral-binding 40-kDa glycoprotein extracted from young adult rabbit alveolar bone.
abstract: A forty-kilodalton (40-kDa) protein was extracted from alveolar bone of young adult rabbit with 0.5 M EDTA after extraction with 4 M GuHCl, and purified by gel-filtration, anion-exchange and hydroxyapatite columns using a high-pressure liquid chromatography system under denaturing conditions. The purified 40-kDa protein was not susceptible to bacterial collagenase and thrombin, but was cleaved by cyanogen bromide. The protein was stained blue with Stains-all. Among various lectins, concanavalin A and lentil lectin agglutinin bound to this protein, but peanut agglutinin, Ricinus communis agglutinin, phytohemagglutinin-E and wheatgerm lectin agglutinin did not. Lectin binding assays showed that the protein is a glycoprotein containing large amounts of mannose and/or glucose residues, but is not a fragment of proteoglycan. The amino acid composition of the protein shows a characteristically high content of acidic amino acids. Therefore, the mineral-binding 40-kDa glycoprotein is considered to be osteonectin/secreted protein acidic and rich in cysteine (SPARC), in terms of similarities to bovine and porcine osteonectins with regard to molecular weight and contents of glycoses and amino acids.
authors.affiliation: Department of Biochemistry, Nihon University School of Dentistry, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Maeno
authors.forename: M
authors.initials: M
authors.lastname: Taguchi
authors.forename: N
authors.initials: N
authors.lastname: Suzuki
authors.forename: T
authors.initials: T
authors.lastname: Ogoshi
authors.forename: K
authors.initials: K
authors.lastname: Naganuma
authors.forename: Y
authors.initials: Y
authors.lastname: Tamura
authors.forename: K
authors.initials: K
authors.lastname: Nakano
authors.forename: F
authors.initials: F
authors.lastname: Komori
authors.forename: K
authors.initials: K
authors.lastname: Otsuka
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
mesh_terms: D000539:Alveolar Process / Q000737:chemistry*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D008334:Mandible / Q000737:chemistry; D003012:Microbial Collagenase; D008903:Minerals / Q000737:chemistry; D008970:Molecular Weight; D015676:Osteonectin / Q000737:chemistry / Q000302:isolation & purification*; D010084:Oxidation-Reduction; D011817:Rabbits; D012685:Sepharose; D012834:Silver; D012967:Sodium Dodecyl Sulfate
chemical_list: D000596:Amino Acids; D008903:Minerals; D015676:Osteonectin; D012967:Sodium Dodecyl Sulfate; D012834:Silver; D012685:Sepharose; D003012:Microbial Collagenase; D003488:Cyanogen Bromide