Pereira B, Wu K K, Wang L H
Department of Internal Medicine, University of Texas-Houston 77030.
Biochem Biophys Res Commun. 1993 Dec 30;197(3):1041-8. doi: 10.1006/bbrc.1993.2583.
Prostacyclin synthase (PGIS) from bovine aorta was purified using conventional purification procedures including detergent solubilization, Sephacryl S-300 gel filtration and Mono Q high performance liquid chromatography. Polyclonal antiserum raised against the 52-kDa protein bound specifically to PGIS. After immobilization to Protein A-sepharose, the PGIS-antibody complex displayed PGIS activity. Results based on (1) elution profile of the Mono Q column, (2) two-dimensional gel electrophoresis and (3) N-terminal amino acid sequence suggested that PGIS is heterogenous. Amino acid sequences of N-terminus and a tryptic peptide led us to isolate a partial cDNA fragment.
采用包括去污剂增溶、Sephacryl S - 300凝胶过滤和Mono Q高效液相色谱在内的传统纯化方法，从牛主动脉中纯化前列环素合酶（PGIS）。针对该52 kDa蛋白产生的多克隆抗血清能特异性结合PGIS。固定于蛋白A - 琼脂糖后，PGIS - 抗体复合物表现出PGIS活性。基于（1）Mono Q柱洗脱图谱、（2）二维凝胶电泳和（3）N端氨基酸序列的结果表明PGIS具有异质性。N端和一个胰蛋白酶肽段的氨基酸序列使我们分离出一个部分cDNA片段。
