Mouse procollagen IV. Characterization and supramolecular association.
The endodermal cell line PF-HR9, derived from the murine teratocarcinoma cell line PCC4-F, was grown as monolayers and as cell clusters called embryoid bodies. Procollagen IV and laminin were isolated from both kinds of culture media. Antibodies specific to collagen IV and to laminin demonstrated these materials in association with the cells and in the culture media. The procollagen IV consisted of pro alpha 1 IV and pro alpha 2 IV chains and gave a circular dichroic spectrum characteristic for collagen helices, with thermal transitions at 40, 44, and 51 degrees C. The molecules were visualized electron microscopically after rotary shadowing. Laminin showed the characteristic beaded cross-appearance, and procollagen IV was a 434 +/- 12-nm long linear thread containing a 17-nm carboxyl-terminal knob. The 7% of collagen helix with Tm = 51 degrees C corresponds to about a 30-nm length of the molecule and is probably that section of the amino end through which several procollagen IV molecules form a junctional complex. Several noncovalent associations of procollagen IV molecules were demonstrated by velocity sedimentation and electron microscopy of concentrated culture media, specifically associations of two and four procollagen IV molecules through their amino ends and dimers linked at their carboxyl ends. The results show that procollagen IV molecules associate noncovalently into the components which others have isolated from basement membranes and strongly support a network model of these supramolecular assemblies.
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D002942:Circular Dichroism; D004707:Endoderm; D006023:Glycoproteins / Q000302:isolation & purification*; D006358:Hot Temperature; D007158:Immunologic Techniques; D007797:Laminin; D046911:Macromolecular Substances; D051379:Mice; D008854:Microscopy, Electron; D009363:Neoplasm Proteins / Q000302:isolation & purification; D011347:Procollagen / Q000302:isolation & purification*; D013724:Teratoma
D006023:Glycoproteins; D007797:Laminin; D046911:Macromolecular Substances; D009363:Neoplasm Proteins; D011347:Procollagen