Type IV collagen from chicken muscular tissues. Isolation and characterization of the pepsin-resistant fragments.
Type IV collagen has been isolated from adult chicken gizzard after limited pepsin digestion and subsequent differential salt fractionation in acidic and neutral conditions. After denaturation, three fragments (called F1, F2, and F3) were isolated by agarose gel filtration and carboxymethylcellulose chromatography. F1 and F2 possessed apparent molecular weights of 53 000 and 50 000, respectively, and were consistently isolated in a 2:1 proportion. F3 was larger and after reduction of disulfide bonds gave rise to three fragments (called F3A, F3B, and F3C) of apparent molecular weights 68 000, 40 000, and 29 000. No alpha-chain-sized components of Type IV collagen were observed. A native fraction containing F1 and F2, but no F3, was isolated after extraction using less pepsin and an additional salt fractionation in acidic conditions. F1 and F2 in the native form were not separated by carboxymethylcellulose or diethylaminoethylcellulose chromatography performed in nondenaturating conditions or by differential salt precipitation in acidic or neutral conditions; these results suggest that F1 and F2 arise as a single native component of structure (F1)2F2. The fraction containing F1 and F2 also gave rise to a single segment long spacing crystallite pattern and to a circular dichroism spectrum which was typical for a native collagen. F1 and F2 were also isolated from chicken heart, blood vessels, and skeletal muscle, whereas from bovine aorta, using the same isolation procedures, two alpha-chain-sized components were obtained, which appeared to be similar to the two Type IV chains recently described by other groups. The data suggest that (i) pepsin fragmentation of type IV collagen from chicken tissues occurs in a different manner compared to Type IV collagen from mammalian tissues and (ii) for the chicken there must be at least two Type IV chains which are assembled into a single native molecule.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002645:Chickens; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003094:Collagen / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005895:Gizzard, Avian / Q000032:analysis; D046911:Macromolecular Substances; D008970:Molecular Weight; D009130:Muscle, Smooth / Q000032:analysis*; D010434:Pepsin A; D010446:Peptide Fragments / Q000032:analysis
D000596:Amino Acids; D046911:Macromolecular Substances; D010446:Peptide Fragments; D003094:Collagen; D010434:Pepsin A