CHARACTERIZATION OF MUSCLE EPIMYSIUM, PERIMYSIUM AND ENDOMYSIUM COLLAGENS
CHARACTERIZATION OF MUSCLE EPIMYSIUM, PERIMYSIUM AND ENDOMYSIUM COLLAGENS
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DOI：
10.1042/bj2191017
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发表时间：
1984-01-01
影响因子：
4.1
通讯作者：
CHAMPION, AE
中科院分区：
文献类型：
--
作者：
LIGHT, N;CHAMPION, AE
由于肌肉中胶原蛋白相对较少，因此难以从肌肉中分离和表征胶原蛋白。这里，提出了一种综合方法，结合了McCollester（1962）和Laurent、Cockerill、McAnglands和Hastings（1981）先前描述的方法，其中[牛]肌肉结缔组织的3个主要部分，即肌外膜、肌束膜和肌内膜，可以从大量肌肉蛋白中制备和分离。由此制备的结缔组织可用盐洗涤并用胃蛋白酶处理以释放可溶性天然胶原，或可用十二烷基硫酸钠洗涤以产生非常干净的不溶性胶原产物。后一种类型的制备可用于定量胶原蛋白的主要遗传形式的比率或用于测量可还原的交联含量以给出可再现的结果。肌外膜和肌束膜都含有I型胶原作为主要成分，III型胶原作为次要成分;肌束膜还含有痕量的V型胶原。肌内膜，个别肌纤维的鞘，被证明含有I型和III型胶原蛋白作为主要成分。V型胶原也存在少量，IV型胶原，基底膜的胶原成分，从肌内膜制剂中纯化。这是第一次在肌肉肌内膜中存在IV型胶原蛋白的生物化学证明。制备是非常相似的其他IV型胶原蛋白从其他基底膜上的十二烷基硫酸钠/聚丙烯酰胺凝胶电泳，是无法区分的EHS肉瘤胶原蛋白和胎盘IV型胶原蛋白的EM旋转阴影后。
It has been difficult to separate and characterize collagen from muscle because of its relative paucity in this tissue. Here, a comprehensive methodology is presented, combining methods previously described by McCollester (1962) and Laurent, Cockerill, McAnulty and Hastings (1981), in which the 3 major tracts of [bovine] muscle connective tissue, the epimysium, perimysium and endomysium, may be prepared and separated from the bulk of muscle protein. Connective tissue thus prepared may be washed with salt and treated with pepsin to liberate soluble native collagen, or can be washed with sodium dodecyl sulfate to produce a very clean insoluble collagenous product. This latter type of preparation may be used for quantification of the ratio of the major genetic forms of collagen or for measurement of reducible cross-link content to give reproducible results. Both the epimysium and perimysium contain type-I collagen as the major component and type-III collagen as a minor component; perimysium also contained traces of type-V collagen. The endomysium, the sheaths of individual muscle fibers, was shown to contain type-I and type-III collagen as major components. Type-V collagens was also present in small amounts, and type-IV collagen, the collagenous component of basement membranes, was purified from endomysial preparations. This is the 1st biochemical demonstration of the presence of type-IV collagen in muscle endomysium. The preparation was very similar to other type-IV collagens from other basement membranes on sodium dodecyl sulfate/polyacrylamide-gel electrophoresis and was indistinguishable from EHS sarcoma collagen and placenta type-IV collagen in the EM after rotary shadowing.