Characterization of a collagenous cementum-derived attachment protein.
We report further characterization of a cementum-derived protein that promotes the adhesion and spreading of periodontal cells. The cementum attachment protein (CAP) was extracted from bovine cementum, separated by diethylamino ethyl (DEAE)-cellulose chromatography, and purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and C18 reverse phase high performance liquid chromatography. The purified preparation contained a single protein band migrating with M(r) 56,000. It did not cross-react with polyclonal antibodies to osteopontin, vitronectin, or other attachment proteins. The attachment activity was resistant to chondroitinase ABC digestion. An internal amino acid sequence of six peptides was determined by microsequencing, and the peptide sequences were not present in other attachment proteins described in cementum. Four sequences contained Gly-X-Y repeats typical of collagen helix. One 17 amino acid peptide had 82% homology with a type XII collagen domain. However, bovine type XII collagen did not promote fibroblast attachment. Although another 19-amino-acid-long peptide had 95% homology to bovine alpha 1 [I], two other peptides were only 74% and 68% homologous, and the CAP was not recognized by anti-type I collagen antibody. The attachment activity of CAP was susceptible to bacterial collagenase. The CAP did not cross-react with antibodies to type V, XII, and XIV collagens. These data and our previous immunostaining data indicate that the CAP is not related to other collagens or attachment proteins and that it is a collagenous attachment protein localized in cementum.
Wu
D
D
Department of Pathology, School of Medicine, University of Washington, Seattle, USA.
Ikezawa
K
K
Parker
T
T
Saito
M
M
Narayanan
A S
AS
D000818:Animals; D000906:Antibodies; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D003094:Collagen / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions; D003739:Dental Cementum / Q000378:metabolism*; D006801:Humans; D010519:Periodontium / Q000378:metabolism / Q000473:pathology*; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology
D000906:Antibodies; D011506:Proteins; D003094:Collagen