Characteristics and localization of rat submandibular gland proteoglycans.
The submandibular gland proteoglycans were investigated biochemically and immunohistochemically in male Sprague-Dawley rats. Proteoglycans were extracted with 4 M guanidine-HCl, followed by ultracentrifugation in a CsCl density gradient, and fractionated by ion-exchange chromatography and gel filtration. The molecular weight of PGs was estimated by SDS-PAGE and immunoblot analysis with monoclonal antibodies (HepSS-1 or 6-B-6). The glycosaminoglycan side-chains in the proteoglycan fractions were identified by electrophoresis on cellulose acetate membrane. Three proteoglycan fractions were obtained. One was a heparan sulphate proteoglycan that migrated as a diffuse band of about 210 kDa. The other two fractions contained at least two dermatan sulphate proteoglycans of 70-85 kDa and 40-50 kDa. Digestion of these two proteoglycans with chondroitinase ABC, but not heparitinase, produced two bands of 50 and 21 kDa, which were core proteins. The smaller dermatan sulphate proteoglycan may be a portion of the other, as the core protein of both bound to 6-B-6 antibody, and sugar chains of both were the same (20-30 kDa). Heparan sulphates recognized by antibody HepSS-1 were observed widely in the basement membrane, fibrous connective tissue, and striated and excretory ductal cells, while dermatan sulphate proteoglycans recognized by antibody 6-B-6 were located in the connective tissue surrounding striated and excretory ducts.
Kamada
A
A
Department of Biochemistry, Osaka Dental University, Japan.
Tamura
I
I
Okazaki
J
J
Matsukawa
F
F
Sakaki
T
T
D000818:Animals; D000911:Antibodies, Monoclonal; D001485:Basement Membrane / Q000737:chemistry; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003238:Connective Tissue / Q000737:chemistry; D003871:Dermatan Sulfate / Q000032:analysis; D004588:Electrophoresis, Cellulose Acetate; D004591:Electrophoresis, Polyacrylamide Gel; D006497:Heparan Sulfate / Q000032:analysis; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008970:Molecular Weight; D011509:Proteoglycans / Q000032:analysis / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018987:Salivary Ducts / Q000737:chemistry; D012471:Salivary Proteins and Peptides / Q000032:analysis / Q000737:chemistry*; D013363:Submandibular Gland / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D003871:Dermatan Sulfate; D006497:Heparan Sulfate; D011509:Proteoglycans; D012471:Salivary Proteins and Peptides