Characteristics of a 28-kDa collagenous protein extracted with guanidine from EDTA-demineralized rabbit alveolar bone.
Bone proteins in alveolar bone of mandibles from young adult rabbits (3-month-old) were extracted with 4.0 M guanidine hydrochloride (GuHCl), followed by 0.5 M ethylenediaminetetraacetate, and again with 4.0 M GuHCl (G2-ext). The proteins in the G2-ext were fractionated on a gel-filtration column, followed by an anion-exchange column in the presence of 7.0 M urea. A 28-kDa protein was isolated from the G2-ext. The purified 28-kDa protein showed intense staining with silver on SDS-PAGE slab-gel under reducing conditions. This protein was digested with bacterial collagenase, and a 19-kDa fragment appeared on the gel. However, the protein was not susceptible to reduction with cyanogen bromide. The protein did not bind to hydroxyapatite crystals in the presence of 7.0 M urea, and also did not bind to some lectins. On SDS-PAGE under non-reducing conditions, the protein migrated as two bands; a new band appeared at approximately the 85-kDa region in addition to the original 28-kDa band. The amino acid compositions of the protein were similar to those of the alpha 1-pN-propeptide of type I procollagen obtained from other tissues.
Maeno
M
M
Department of Biochemistry, Nihon University School of Dentistry, Tokyo, Japan.
Oikawa
E
E
Taguchi
M
M
Suzuki
N
N
Shiraishi
Y
Y
Hayashi
A
A
Yahagi
N
N
Katayama
I
I
Otsuka
K
K
D000539:Alveolar Process / Q000737:chemistry*; D000818:Animals; D003094:Collagen / Q000302:isolation & purification*; D008334:Mandible / Q000737:chemistry; D010748:Phosphopeptides / Q000302:isolation & purification*; D011347:Procollagen / Q000302:isolation & purification*; D011817:Rabbits
C062121:N-propeptide type I collagen; D010748:Phosphopeptides; D011347:Procollagen; D003094:Collagen