Characterization of collagens of diseased human gingiva.

Abstract

In the gingiva and other connective tissues, alteration in the collagens is primarily responsible for their functional impairment during disease. To study the collagen alterations, we extracted diseased human gingival tissue with neutral and acidic solvents and then with pepsin. The pepsin extract was separated into proteins soluble in 2.5 and 1.5 M NaCl and proteins insoluble in 1.5 M NaCl. By the criteria of solubility behavior in NaCl solutions, elution from (carboxymethyl)cellulose (CM-cellulose) columns, sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis, CNBr peptide pattern, and amino acid composition, the collagens of acidic and neutral solvent extracts and 1.5 M soluble fraction of pepsin extract were characterized as type I collagen and the 1.5 M NaCl insoluble collagen as type III. The 2.5 M NaCl fraction contained alpha 2, A, and B chains. The alpha 1 chains resembled alpha 1[I] in amino acid composition, and, since alpha 2 chains were lacking, it appeared that these chains derived from type I trimer collagen. The A and B chains were purified from the 2.5 M NaCl fractions by salting out at acidic pH. The final (A plus B) chain fraction was resolved into two major and one minor protein peaks by phosphocellulose chromatography. The major peaks were characterized as A and B chains on the basis of amino acid composition and CNBr peptide patterns. The minor peak had electrophoretic mobility slightly less than B chains, and the amino acid composition was different. Analysis of the proportion of different collagen types extracted indicated that type III collagen, which is the second major fraction in other connective tissues, is only a minor constituent in the gingiva. More interestingly, A and B chains accounted for a greater proportion than type III. Unlike the fibroblast cultures, the type I trimer formed only a small proportion of collagens of diseased gingival tissue.