p-HMW-collagen, a minor collagen obtained from chick embryo cartilage without proteolytic treatment of the tissue.
p-HMW-collagen, a minor collagen obtained from chick embryo cartilage without proteolytic treatment of the tissue.
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p-HMW-胶原蛋白，一种从鸡胚软骨中获得的少量胶原蛋白，未经组织蛋白水解处理。
DOI：
10.1111/j.1432-1033.1983.tb07746.x
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Tuderman,L
中科院分区：
文献类型：
--
作者：
Bruckner,P;Mayne,R;Tuderman,L
在胃蛋白酶处理幼龄鸡的胸骨软骨后，分离出软骨的次要胶原蛋白片段（称为HMW和LMW），并通过其圆二色性光谱判断为完全为三螺旋分子。对HMW的复性动力学研究表明，HMW的链间二硫键位于长臂的一端。这些抗体与II型胶原及其他次要胶原如天然或变性结构的LMW和1α、2α、3α胶原均无交叉反应，并用于鉴定鸡胚软骨体外合成的HMW相关分子。这些分子中的一些被分泌到器官培养基中，并且可以通过硫酸铵沉淀从器官培养基中回收。该沉淀物的聚丙烯酰胺凝胶电泳得到一条高分子量条带，当通过免疫印迹鉴定时，该条带可减少为迁移速度略快于α1（II）链的两条条带。当通过聚丙烯酰胺凝胶电泳和荧光照相法分析时，也可以在培养基蛋白的硫酸铵沉淀物中存在的约6种放射性标记多肽中鉴定出这些条带，可以通过抗HMW抗体免疫沉淀从培养基中回收相同的多肽。通过从软骨组织中提取并在聚丙烯酰胺凝胶上分离的材料的免疫印迹显示它们在软骨组织中的存在。我们认为，含有这些多肽链的蛋白质代表了体内合成的消化片段HMW的母体分子，并将其命名为p-HMW-胶原。
The fragments of minor collagens of cartilages, called HMW and LMW, were isolated after pepsin treatment of sternal cartilages of young chickens and were shown to be entirely triple‐helical molecules as judged by their circular dichroic spectra. Studies on renaturation kinetics of HMW suggested that the interchain disulfide bonds in HMW reside at one of the ends of the so‐called long arm.Polyclonal antibodies against HMW were raised and affinity purified. These antibodies did not cross‐react with type II collagen nor with other minor collagens such as LMW and 1α, 2α, 3α collagen in native or denatured structure.The antibodies were used to identify HMW‐related molecules which were synthesized by embryonic chick cartilagesin vitro. Some of these molecules were secreted into the organ culture medium and could be recovered from it by ammonium sulfate precipitation. Polyacrylamide gel electrophoresis of this precipitate gave one band of high molecular weight which could be reduced to two bands migrating slightly faster than the α1(II) chain when identified by immunoblotting. These bands could also be identified among about six radiolabelled polypeptides present in the ammonium sulfate precipitate of medium proteins when analysed by polyacrylamide gel electrophoresis followed by fluorography.The same polypeptides could be recovered from the medium by immunoprecipitation with anti‐HMW antibodies. Their presence in cartilage tissue was shown by immunoblotting of material extracted from cartilage tissue and separated on polyacrylamide gels. We suggest that the protein containing these polypeptide chains represents the parent molecule of the peptic fragment HMW as it is synthesizedin vivoand have designated it p‐HMW‐collagen.