Procollagen is more stable in cellulo than in vitro.
The thermal denaturation of both intracellular and freshly secreted chick embryo tendon type I procollagen was investigated using susceptibility to proteolysis by trypsin and chymotrypsin as a probe for triple-helical conformation. Freshly secreted procollagen from the medium of matrix-free tendon cells in suspension or procollagen within the cells and in the pericellular environment melted at 45 degrees C. In contrast, if freshly secreted procollagen was subjected to the melting procedure after dialysis of the medium against 0.4 M NaCl, 0.1 M Tris HCl, pH 7.4 the protein melted at 42 degrees C, the melting temperature of purified procollagen dissolved in the same buffer. In each of these cases, the thermal denaturation profile was narrow, with a width of 1.0-1.5 degrees C. These results demonstrate that, in situ, procollagen is more stable toward thermal denaturation than was previously thought. This extra margin of thermal stability partially resolves the dilemma of how tissues are able to assemble triple-helical procollagen molecules at body temperatures that closely approach the melting temperature of the purified protein.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism; D005470:Fluorometry; D011347:Procollagen / Q000378:metabolism*; D011489:Protein Denaturation; D013710:Tendons / Q000378:metabolism; D013816:Thermodynamics
D011347:Procollagen