Collagen fibrillogenesis in vitro: interaction of types I and V collagen regulates fibril diameter.
Collagen fibrillogenesis in vitro: interaction of types I and V collagen regulates fibril diameter.
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DOI：
10.1017/s0424820100142670
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发表时间：
1990-04
影响因子：
4
通讯作者：
D. Birk;J. Fitch;J. Babiarz;K. Doane;T. Linsenmayer
中科院分区：
文献类型：
--
作者：
D. Birk;J. Fitch;J. Babiarz;K. Doane;T. Linsenmayer
鸡角膜基质的小直径原纤维是异型的，由 I 型和 V 型胶原组成。相对于其他具有较大直径原纤维的含有 I 型的组织，该组织具有高浓度的 V 型胶原，表明异型相互作用可能在控制原纤维直径方面具有调节作用。使用体外自组装系统研究了 I 型和 V 型胶原蛋白的相互作用。在蛋白酶抑制剂存在下，从山黧鸡胚胎中纯化胶原蛋白。 V型胶原制剂含有较高分子量形式的α1(V)和α2(V)链，占总量的60-70%。旋转阴影电子显微照片显示，存在一个小的胃蛋白酶敏感末端区域，其数量与电泳所见一致。在体外，这种纯化的V型胶原蛋白形成了没有明显周期性的细原纤维，而I型胶原原纤维则具有大直径的广泛分布。然而，当 I 型胶原与越来越多的 V 型胶原混合时，观察到 D 周期性原纤维的平均原纤维直径和方差逐渐显着减小。 V 型胶原蛋白分子的氨基末端结构域是这种调节作用所必需的，并且在缺乏氨基末端结构域的情况下，几乎观察不到直径减小活性。使用胶原蛋白类型特异性单克隆抗体的电子显微镜表明，形成的原纤维是异型的，含有 I 型和 V 型胶原蛋白。这些数据表明 V 型与 I 型胶原蛋白的相互作用是调节原纤维直径的一种机制，并且至少部分负责调节胶原原纤维形成。
The small-diameter fibrils of the chick corneal stroma are heterotypic, composed of both collagen types I and V. This tissue has a high concentration of type V collagen relative to other type I-containing tissues with larger-diameter fibrils, suggesting that heterotypic interactions may have a regulatory role in the control of fibril diameter. The interactions of collagen types I and V were studied using an in vitro self-assembly system. Collagens were purified from lathyritic chick embryos in the presence of protease inhibitors. The type V collagen preparations contained higher molecular weight forms of the alpha 1(V) and alpha 2(V) chains constituting 60-70% of the total. Rotary-shadow electron micrographs showed a persistence of a small, pepsin-sensitive terminal region in an amount consistent with that seen by electrophoresis. In vitro, this purified type V collagen formed thin fibrils with no apparent periodicity, while type I collagen fibrils had a broad distribution of large diameters. However, when type I collagen was mixed with increasing amounts of type V collagen a progressive and significant decrease in both the mean fibril diameter and the variance was observed for D periodic fibrils. The amino-terminal domain of the type V collagen molecule was required for this regulatory effect and in its absence little diameter reducing activity was observed. Electron microscopy using collagen type-specific monoclonal antibodies demonstrated that the fibrils formed were heterotypic, containing both collagen types I and V. These data indicate that the interaction of type V with type I collagen is one mechanism modulating fibril diameter and is at least partially responsible for the regulation of collagen fibril formation.