Assembly of chick and bovine lens-capsule collagen.
Chick-embryo and adult bovine lens-capsular epithelia in organ culture synthesized 4-hydroxy[3H]proline-containing polypeptides when incubated in the presence of [3H]proline. These collagenous polypeptides of apparent Mr 180 000, 175 000 and 160 000 became incorporated with time into aggregates of higher molecular size. The formation of such aggregates was inhibited when the tissues were labelled in the presence of beta-aminopropionitrile, thereby implicating lysine-derived cross-links in aggregate formation. When the tissues were incubated in the presence of tunicamycin, the collagenous polypeptides synthesized exhibited increased electrophoretic mobilities on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The addition to lens-capsule incubation medium of alpha alpha'-bipyridine led to the synthesis of underhydroxylated type IV collagen, also of increased electrophoretic mobility. Extended pulse-chase experiments indicated that such underhydroxylated collagen did not participate in aggregate formation, but was at least as stable as fully hydroxylated non-cross-linked collagen synthesized in the presence of beta-aminopropionitrile. Native type IV collagen, recovered from the culture medium when capsules were incubated with [3H]proline for 24h, was purified by ion-exchange chromatography. Separations conducted on CM-cellulose under denaturing and nondenaturing conditions suggested that the alpha 1(IV) and alpha 2(IV) chains occur in the same heterologous triple helix. Densitometric analyses of appropriate fluorograms indicated that these two polypeptides occur in a 2:1 ratio, suggesting that lens-capsule collagen is synthesized as a triple-helical molecule of composition [alpha 1(IV)]2 alpha 2(IV).
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D015082:2,2'-Dipyridyl / Q000494:pharmacology; D000629:Aminopropionitrile / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002645:Chickens; D002852:Chromatography, Ion Exchange; D002918:Chymotrypsin; D003094:Collagen / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D007903:Lens Capsule, Crystalline / Q000378:metabolism*; D007908:Lens, Crystalline / Q000378:metabolism*; D003012:Microbial Collagenase; D010434:Pepsin A; D010446:Peptide Fragments / Q000032:analysis; D011392:Proline / Q000378:metabolism; D013045:Species Specificity; D014415:Tunicamycin / Q000494:pharmacology
D010446:Peptide Fragments; D014415:Tunicamycin; D000629:Aminopropionitrile; D015082:2,2'-Dipyridyl; D003094:Collagen; D011392:Proline; D002918:Chymotrypsin; D010434:Pepsin A; D003012:Microbial Collagenase