ISOLATION AND PURIFICATION OF PROSTAGLANDIN E ISOMERASE

前列腺素E异构酶(也称为PGE2合成酶)催化 前列腺素内源性过氧化产物(PGH2)的异构化反应 已转换为PGE2。PGE2的强大生物活性表明 前列腺素E异构酶在卵巢癌发病机制中的重要作用 牙周病、肿瘤进展、炎症和免疫 回应。一种精制制剂的商业应用 因此哺乳动物的前列腺素E异构酶应该促进碱性和 药物研究，包括对参与的因素的研究 前列腺素E异构酶基因表达的调控，新的研究进展 治疗癌症、糖尿病和癌症的药物 某些形式的精神分裂症的病因。 在第一阶段，我们将从公羊精液中纯化前列腺素E异构酶 小泡(RSV)，确定酶测定和储存的条件，以及 评估纯化蛋白的辅因子需求量。在第二阶段 研究，我们将使用亲和层析和免疫亲和 用层析法将酶提纯到均一。纯化的酶 将用于生产多克隆抗体和氨基酸 测序。基于多克隆抗体和寡核苷酸探针的研究 部分氨基酸序列将被制备并用作试剂 前列腺素分子克隆的鉴定与分离 E异构酶基因及其进一步鉴定。

Prostaglandin E isomerase (known as PGE2 synthase also) catalyses the isomerization reaction in which prostaglandin endoperoxide (PGH2), is converted to PGE2. The potent biological activities of PGE2 indicate an important role for prostaglandin E isomerase in the pathogenesis of periodontal disease, progression of the tumor, inflammatory and immune responses. Commercial availability of a purified preparation of mammalian prostaglandin E isomerase should therefore facilitate basic and pharmaceutical research, including studies of factors involved in regulation of PGE isomerase gene expression, development of novel therapeutic agents for the treatment of cancer, diabetes, and the pathogeny of certain forms of schizophrenia. In Phase I, we will purify prostaglandin E isomerase from ram seminal vesicles (RSV), determine conditions for enzyme assay and storage, and evaluate the cofactor requirements of the purified protein. In Phase II research, we will employ affinity chromatography and immunoaffinity chromatography to purify the enzyme to homogeneity. The purified enzyme will be used for the production of polyclonal antibodies and amino acid sequencing. Polyclonal antibodies and oligonucleotide probes based on a partial amino acid sequence will be prepared and used as reagents for the identification and isolation of molecular clones of the prostaglandin E isomerase gene and its further characterization.