Timpl R, Glanville R W, Nowack H, Wiedemann H, Fietzek P P, Kühn K
Hoppe Seylers Z Physiol Chem. 1975 Nov;356(11):1783-92. doi: 10.1515/bchm2.1975.356.2.1783.
Extraction of fetal bovine skin at neutral pH and in the presence of protease inhibitors solubilized substantial amounts of type I and type type III collagen and Type III procollagen. Type I and Type III collagen were separated from each other by salt precipitation, and DEAE-cellulose chromatography was used to separate collagen from procollagen. The main chain constituents in type III collagen and procollagen were disulfide-bonded gamma and pgamma components, respectively. Amino acid composition, cross striation banding as observed using electron microscopy, cyanogen bromide peptide patterns in disc electrophoresis gels and resistance of the disulfide regions to pepsin digestion indicated a close similarity to previously described insoluble type III collagen, which was solubilized by limited pepsin digestion. Electron microscopy of long-spacing-segment crystallites and evidence for an extended form of the disulfide-bonded cyanogen bromide peptide suggested that neutral-salt-soluble type III collagon is longer at its C-terminal end by about 10 to 20 amino acid residues than pepsin-treated material. A small elongation was also indicated in the N-terminal portion of the molecule. Procollagen has an additional N-terminal extension with a length of about 160 A, but no difference was observed between collagen and procollagen at their C-terminal ends.
在中性pH值且存在蛋白酶抑制剂的条件下提取胎牛皮肤，可溶解大量的I型和III型胶原蛋白以及III型前胶原。通过盐沉淀法将I型和III型胶原蛋白彼此分离，并用DEAE - 纤维素色谱法将胶原蛋白与前胶原分离。III型胶原蛋白和前胶原中的主链成分分别是通过二硫键连接的γ和pgamma组分。氨基酸组成、电子显微镜观察到的横纹带、圆盘电泳凝胶中的溴化氰肽图谱以及二硫键区域对胃蛋白酶消化的抗性表明，其与先前描述的经有限胃蛋白酶消化可溶解的不溶性III型胶原蛋白非常相似。长间距片段微晶的电子显微镜观察以及二硫键连接的溴化氰肽的延伸形式的证据表明，中性盐溶性III型胶原蛋白在其C末端比经胃蛋白酶处理的材料长约10至20个氨基酸残基。在分子的N末端部分也显示出小的延伸。前胶原具有约160 Å的额外N末端延伸，但在胶原蛋白和前胶原的C末端未观察到差异。
