Ultrastructural morphology and domain structure of a unique collagenous component of basement membranes.
Ultrastructural morphology and domain structure of a unique collagenous component of basement membranes.
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基底膜独特胶原成分的超微结构形态和域结构。
DOI：
10.1021/bi00281a005
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发表时间：
1983
期刊：
影响因子：
2.9
通讯作者：
Furthmayr,H
中科院分区：
文献类型：
--
作者：
Madri,JA;Foellmer,HG;Furthmayr,H
材料与方法：胶原蛋白制剂。如前所述，从小鼠Engelbreth-Holm-Swarm肿瘤基质中制备酸溶性IV型胶原（Roll et al., 1980）。I型、III型、IV型和V型胶原是由先前描述的人胎盘绒毛、胎盘膜和牛肺实质制备的（Madri & Furthmayr, 1980; Foellmer et al., 1982）。7S片段的分离是根据对risteeli等人（1980）和Furuto & Miller（1980, 1981）方法的改进进行的，如图1所示。短暂、组织(新鲜小牛肺从当地屠宰场,获得人类胎盘从交付套件,获得或肿瘤小鼠EHS矩阵)剁碎,水洗,并与胃蛋白酶消化(沃辛顿):酶底物的比例1:10(干重/干重)dilutedacetic酸(0.001米)的24小时收集4 c合成supemate bycentrifugation,和固体氯化钠添加到0.7 M .第三我和胶原蛋白的合成precipitatecontains类型。将固体氯化钠加入到超络合物中，最终浓度为1.2 m，形成的沉淀物含有IV型和V型胶原蛋白。然后将超消旋物与氯化钠作1.8 M的反应，形成的沉淀对0进行透析。001 M醋酸。在1.8 M溶液中加入固体氯化钠再沉淀后，将颗粒溶解，用稀乙酸进行透析
Materials and MethodsCollagen Preparations. Acid-soluble type IV collagen was prepared from the murine Engelbreth-Holm-Swarm tumor matrix as previously described (Roll et al., 1980). Types I, III, IV, and V collagens were prepared from human placental villi and placental membranes and bovinelung parenchyma as previously described (Madri & Furthmayr, 1980; Foellmer et al., 1982). Isolation of the 7S fragment was performed according to a modification of the methods of Risteli et al.(1980) and Furuto & Miller (1980, 1981) as outlined in Figure 1. Briefly, tissue (fresh calf lung obtained from a local slaughterhouse, human placentas obtained from the delivery suite, or murine EHS tumor matrix) is minced, washed, and digested with pepsin (Worthington) at an enzyme: substrate ratio of 1: 10 (dry weight/dry weight) in dilutedacetic acid (0.001 M) for 24 h at 4 C. The resultant supemate is collected bycentrifugation, and solid sodium chloride is added to 0.7 M. The resultant precipitatecontains types I and III collagen. Solid sodium chloride is added to the supemate to a final concentration of 1.2 M. The precipitate formed con-tains types IV and V collagen. The supemate is then made 1.8 M with respect to sodium chloride, and the precipitate formed is dialyzed against0. 001 M acetic acid. After being reprecipitated by addition of solid sodium chloride to 1.8 M, the pellet is dissolved and dialyzed against dilute acetic acid