Isolation of Noncollagenous Proteins from Gingival Connective Tissue

Abstract

Noncollagenous proteins form an integral part of gingiva and other connective tissues. We have performed studies aimed at purification and partial characterization of the gingival noncollagenous proteins. Healthy gingival tissues from mongrel dogs were extracted in neutral buffers, acetic acid, and 6 mol/L urea. Immunoblots using anti-keratin antibodies and CNBr peptide patterns revealed that the majority of the proteins present in these extracts were keratins. To exclude keratins, gingival connective tissue was separated from the epithelium and then extracted. Acid extracts of the connective tissue contained very little protein, whereas urea extracts contained collagen and other noncollagenous proteins. The noncollagenous proteins present in the urea extract were partially purified by DEAE-cellulose chromatography and separated by affinity chromatography through a Sepharose 4B-type I collagen column. At least eight proteins, which ranged in molecular size from 15 to 75 kilodaltons, were obtained by this procedure. We conclude that keratins are major components of whole gingiva extracts and that epithelium must first be removed in order for connective tissue proteins to be obtained. The gingival connective tissue appears to contain several collagen-binding proteins, and these proteins may play an important role in the structure and function of the gingival matrix.

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