Characterization of collagen synthesized by normal and chemically transformed rat liver epithelial cell lines.

A rat liver epithelial cell line (K16) and a 2-[N-(acetoxyacetyl)amino]fluorine-transformed K16 cell line (W8) [Weinstein, I.B., Yamaguchi, R., Gerbert, R., & Kaighn, M.E. (1975) In Vitro 11, 130-141] synthesize and secrete similar quantities of collagenous proteins. The transformed cells produce more total protein so that the percent collagen produced is decreased compared to that of the parent cell line. The type of collagen produced by the transformants differed from that of the parent cell line. After limited pepsin digestion, the pepsin-resistant collagen molecules synthesized by K16 cells precipitated at 2.6 M NaCl and contained both alpha 1 (I) and alpha 2 chains in a 2.5:1 ratio. The collagen synthesized by the W8 cells precipitated at higher salt concentrations, and no pepsin-resistant alpha 2 chains could be demonstrated by CM-cellulose chromatography or gel electrophoresis. The cyanogen bromide peptides of the W8 alpha 1 chains did not cochromatograph with type I rat skin collagen cyanogen bromide peptides on CM-cellulose chromatography, whereas the alpha 1 chains of the K16 cells did cochromatograph. The conversion of procollagen to collagen was also decreased in the transformant cells. Therefore, the chemical transformation of K16 produced transformants with increased protein synthesis and altered collagen metabolism.