The specific detection of collagenous proteins after electrophoresis using enzyme-conjugated collagen-binding fibronectin fragments.
The specific detection of collagenous proteins after electrophoresis using enzyme-conjugated collagen-binding fibronectin fragments.
复制标题
使用酶联胶原蛋白结合纤连蛋白片段进行电泳后胶原蛋白的特异性检测。
DOI：
10.1016/0003-2697(82)90454-7
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发表时间：
1982
影响因子：
2.9
通讯作者：
Engvall,E
中科院分区：
文献类型：
--
作者：
Bell,ML;Engvall,E
设计了一种检测通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的蛋白质混合物中的胶原蛋白的方法。将凝胶上的蛋白质转移到硝酸纤维素滤膜上。通过与辣根过氧化物酶缀合物一起孵育，然后为酶促反应提供底物和色原，将它们在滤纸上染色。使用纤连蛋白的胰凝乳蛋白酶片段制备了一种新型酶缀合物，该片段已被证明含有胶原蛋白结合位点。该缀合物用于对 I、II、III、IV 和 V 型胃蛋白酶化人胶原蛋白的多肽链进行染色。它还对胶原蛋白的一些溴化氰片段和补体亚成分 C1q 的所有三个多肽链进行染色。该程序可用于检测异质混合物中的胶原蛋白以及胶原蛋白和胶原蛋白样蛋白的鉴定和表征。
A method to detect collagenous proteins within a mixture of proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis has been devised. Proteins on the gels were transferred to nitrocellulose filters. They were stained on the filters by incubating with horseradish peroxidase conjugates and then providing a substrate and chromogen for the enzymatic reaction. A novel enzyme conjugate was prepared using a chymotryptic fragment of fibronectin which has been shown to contain the collagen-binding site. The conjugate was used to stain polypeptide chains of pepsinized human collagens of types I, II, III, IV, and V. It also stained some cyanogen bromide fragments of collagens and all three polypeptide chains of the complement subcomponent C1q. This procedure will be useful in the detection of collagenous proteins in heterogeneous mixtures and in the identification and characterization of collagens and collagenlike proteins.