短杆菌肽S合成酶。多酶的电泳特性分析。

Gramicidin S-synthetase. Electrophoretic characterization of the multienzyme.

Koischwitz H, Kleinkauf H

Biochim Biophys Acta. 1976 May 13;429(3):1052-61. doi: 10.1016/0005-2744(76)90350-8.

Abstract

A method characterizing the fully active gramicidin S-synthetase (EC. 6.3.2.-) multienzyme in protein mixtures by a combination of sedimentation and polyacrylamide gel electrophoretic mobility data has been described. 2. The molecular weight of 280000 has been reevaluated by gradient centrifugation, gel filtration, and polyacrylamide gel electrophoresis in presence of sodium dodecyl sulfate. The size of the multienzyme is not changed by sodium dodecyl sulfate treatment. 3. In polyacrylamide gel electrophoresis dimerisation occurs in Tris, while two bands, which may represent monomer and dimer, are observed in phosphate. 4. Reliability of molecular weight determinations of sodium dodecyl sulfate-protein complexes of sizes up to 300000 daltons has been determined, correlating either mobilities or retardation coefficients.

摘要

已描述了一种通过沉降和聚丙烯酰胺凝胶电泳迁移率数据相结合的方法，来表征蛋白质混合物中完全活性的短杆菌肽S合成酶（EC. 6.3.2.-）多酶。2. 通过梯度离心、凝胶过滤和在十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳，对280000的分子量进行了重新评估。十二烷基硫酸钠处理不会改变多酶的大小。3. 在聚丙烯酰胺凝胶电泳中，二聚化发生在Tris中，而在磷酸盐中观察到两条带，这两条带可能代表单体和二聚体。4. 已确定了分子量高达300000道尔顿的十二烷基硫酸钠 - 蛋白质复合物分子量测定的可靠性，该可靠性与迁移率或阻滞系数相关。