PHOSPHATIDYLINOSITOL BIOSYNTHESIS IN SACCHAROMYCES-CEREVISIAE - PURIFICATION AND PROPERTIES OF MICROSOME-ASSOCIATED PHOSPHATIDYLINOSITOL SYNTHASE
PHOSPHATIDYLINOSITOL BIOSYNTHESIS IN SACCHAROMYCES-CEREVISIAE - PURIFICATION AND PROPERTIES OF MICROSOME-ASSOCIATED PHOSPHATIDYLINOSITOL SYNTHASE
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DOI：
10.1128/jb.154.1.304-311.1983
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发表时间：
1983-01-01
影响因子：
3.2
通讯作者：
CARMAN, GM
中科院分区：
文献类型：
--
作者：
FISCHL, AS;CARMAN, GM
膜相关磷脂生物合成酶磷脂酰肌醇合酶（胞苷5“-二磷酸-1，2-二酰基-sn-甘油：肌醇3-磷脂酰转移酶，EC 2.7.8.11）从S.啤酒。纯化过程包括TritonX-100溶解微粒体膜，CDP二酰甘油-Sepharose亲和层析和层析聚焦。该程序导致在分离的一个表观最小亚基分子量为34，000，在十二烷基硫酸钠的存在下，通过聚丙烯酰胺凝胶电泳测定的一个几乎均匀的蛋白质制剂。磷脂酰肌醇合酶依赖于锰和Triton X-100的最大活性。最适pH为8.0。硫活性剂抑制酶活性。活化能为35 kcal/mol（146，540 J/mol）。该酶在高达60 ℃的温度下相当稳定。C.
The membrane-associated phospholipid biosynthetic enzyme phosphatidylinositol synthase (cytidine 5''-diphospho-1,2-diacyl-sn-glycerol:myo-inositol 3-phosphatidyltransferase, EC 2.7.8.11) was purified 1000-fold from the microsomal fraction of S. cerevisiae. The purification procedure included Triton X-100 solubilization of the microsomal membranes, CDPdiacylglycerol-Sepharose affinity chromatography and chromatofocusing. The procedure resulted in the isolation of a nearly homogeneous protein preparation with an apparent minimum subunit MW of 34,000, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Phosphatidylinositol synthase was dependent on Mn and Triton X-100 for maximum activity. The pH optimum was 8.0. Thioreactive agents inhibited enzyme activity. The energy of activation was 35 kcal/mol (146,540 J/mol). The enzyme was reasonably stable at temperatures of up to 60.degree. C.