Structure and biosynthesis of rabbit lens capsule collagen.
The present study was designed to compare collagen synthesized by rabbit lens epithelial cells in culture with rabbit lens capsule collagen. Confluent monolayers of rabbit lens epithelial cells were established. Incorporation of [3H]-proline into glycoproteins secreted into the medium and cell surface components were analyzed in the presence of protease inhibitors. Gel filtration chromatography on sodium dodecyl sulfate--agarose (Bio-gel A-5m) of [3H]-labeled newly synthesized proteins by lens epithelial cells in culture resolved into a single precursor of approximate molecular weight of 160,000 daltons. Neither the medium nor the cell layer showed any evidence of low-molecular weight hydroxyproline-containing material. Limited pepsin digestion of this material cleaved the higher molecular weight chains into smaller components ranging from 25,000 to 110,000 daltons. Pepsin digestion and direct extraction of the collagenous components of the rabbit lens capsule revealed materials of high--molecular weight proteins similar to that synthesized in culture. Low--molecular weight (55,000 daltons) protein was only detected in lens capsules after prolonged pepsin digestion. S-Carboxylation of the lens capsules collagens did not affect their mobilities, but repepsinization gave rise to 110,000 dalton protein, although no significant changes in the amino acid composition were noticed. The absence of synthesis of low--molecular weight protein by cell culture and the presence of low--molecular weight components only after prolonged pepsin digestion of lens capsule could be the result of unusual susceptibility of the basement membrane collagens to pepsin attack.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D002850:Chromatography, Gel; D003094:Collagen / Q000096:biosynthesis* / Q000302:isolation & purification; D005136:Eye Proteins / Q000032:analysis; D007903:Lens Capsule, Crystalline / Q000378:metabolism*; D007908:Lens, Crystalline / Q000378:metabolism*; D010434:Pepsin A / Q000494:pharmacology; D011817:Rabbits
D000596:Amino Acids; D005136:Eye Proteins; D003094:Collagen; D010434:Pepsin A