Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein.
Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein.
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椎间盘的 VI 型胶原蛋白。
DOI：
10.1042/bj2480373
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Slayter,HS
中科院分区：
文献类型：
--
作者：
Wu,JJ;Eyre,DR;Slayter,HS
椎间盘的胶原蛋白框架含有两种主要的纤维形成胶原蛋白，I 型和 II 型。还存在少量其他类型的胶原蛋白。在检查牛椎间盘组织内这些次要胶原蛋白的性质和分布时，发现 VI 型胶原蛋白异常丰富。其在小腿髓核中约占胶原蛋白总量的20%，在纤维环中约占胶原蛋白总量的5%。通过用软骨素ABC裂解酶和链霉菌透明质酸酶连续消化椎间盘组织，发现可以提取VI型胶原蛋白的天然共价聚合物。通过旋转阴影制备的这种材料的电子显微照片揭示了 VI 型分子的四聚体和双四聚体的特征尺寸，以及它们的中心杆和末端球状结构域。在非还原非变性条件下在琼脂糖上进行分子筛柱色谱得到一系列蛋白质峰，其分子大小相当于四聚体、双四聚体和更高的多聚体。在二硫化物裂解后的 SDS/聚丙烯酰胺凝胶电泳上，这些 VI 型胶原蛋白级分均显示出位于 Mr 140,000 处的主条带和位于 Mr 180,000 至 240,000 之间的四个次要条带。在琼脂糖/2.4%聚丙烯酰胺中进行无二硫键裂解的电泳时，仅存在VI型分子的二聚体（6链）和四聚体（12链）形式。在 4 M-氯化胍中提取组织的所有 VI 型胶原的能力，以及直接分析中不存在醛介导的交联残基，表明与大多数基质胶原相反，VI 型胶原不能充当共价交联的结构聚合物。
The collagen framework of the intervertebral disc contains two major fibril-forming collagens, types I and II. Smaller amounts of other types of collagen are also present. On examination of the nature and distribution of these minor collagens within bovine disc tissue, type VI collagen was found to be unusually abundant. It accounted for about 20% of the total collagen in calf nucleus pulposus, and about 5% in the annulus fibrosus. It was discovered by serially digesting disc tissue with chondroitin ABC lyase and Streptomyces hyaluronidase that native covalent polymers of type VI collagen could be extracted. Electron micrographs of this material prepared by rotary shadowing revealed the characteristic dimensions of tetramers and double tetramers of type VI molecules, with their central rods and terminal globular domains. Molecular-sieve column chromatography on agarose under non-reducing non-denaturing conditions gave a series of protein peaks with molecular sizes equivalent to the tetramer, double tetramer and higher multimers. On SDS/polyacrylamide-gel electrophoresis after disulphide cleavage, these fractions of type VI collagen all showed a main band at Mr 140,000 and four lesser bands between Mr 180,000 and 240,000. On electrophoresis without disulphide cleavage in agarose/2.4% polyacrylamide only dimeric (six chains) and tetrameric (12 chains) forms of type VI molecules were present. The ability to extract all the type VI collagen of the tissue in 4 M-guanidinium chloride, and absence of aldehyde-mediated cross-linking residues on direct analysis, showed that, in contrast with most matrix collagens, type VI collagen does not function as a covalently cross-linked structural polymer.