通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳鉴定从大鼠肝细胞膜中纯化的磷脂酸磷酸水解酶。

Identification of phosphatidate phosphohydrolase purified from rat liver membranes on SDS-polyacrylamide gel electrophoresis.

作者信息

Siess E A, Hofstetter M M

机构信息

Medizinische Poliklinik der Ludwig-Maximilians-Universität München, Germany.

出版信息

FEBS Lett. 1996 Mar 4;381(3):169-73. doi: 10.1016/0014-5793(96)00111-1.

DOI:10.1016/0014-5793(96)00111-1

PMID:8601448

Abstract

Phosphatidate phosphohydrolase (PAP; EC 3.1.3.4) insensitive to N-ethylmaleimide was partially purified from rat liver membranes by a combination of chromatographic methods, immunoabsorption and glycerol gradient centrifugation. The specific activity was increased more than 600-fold over that of the membrane extract. Enzyme antibodies precipitating more than 80% of PAP were obtained and used for the identification of PAP protein on SDS-polyacrylamide gels employing the immunodetection method of Muilerman et al. [(1982) Anal. Biochem. 120, 46-51]. By this approach PAP was localized as a 31 kDa polypeptide.

摘要

对N-乙基马来酰亚胺不敏感的磷脂酸磷酸水解酶（PAP；EC 3.1.3.4）通过色谱法、免疫吸附和甘油梯度离心相结合的方法从大鼠肝细胞膜中部分纯化。比活性比膜提取物提高了600多倍。获得了沉淀超过80% PAP的酶抗体，并采用Muilerman等人的免疫检测方法[(1982) Anal. Biochem. 120, 46 - 51]在SDS-聚丙烯酰胺凝胶上鉴定PAP蛋白。通过这种方法，PAP被定位为一种31 kDa的多肽。