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出 处:《生物化学杂志》1995年第1期16-21,共6页
摘 要:在纯化猪肝微粒体磷脂酰肌醇-4-激酶(PI-4-K)的基础上,制备该酶的免疫血清、提纯IgG,并建立了PI4-K的ELISA,进行不同组织中PI4-K的免疫沉淀分析和免疫定量。结果证明:猪肝微粒体PI4-K的抗血清或抗体IgG可沉淀猪肾和猪肺的微粒体膜、猪肝细胞膜和人中性粒细胞细胞膜TritonX-100增溶液(TXSM)中的PI4-K,表明不同组织、不同亚细胞器和不同种属的PI4-K有相似的抗原性。猪肝、肾、肺、胃、小肠和大肠TXSM中PI4-K的活力和酶蛋白含量相仿,急性中性粒细胞白血病患者的中性粒细胞膜上PI4-K的含量和活力都增加3.5倍左右.加上PI4-K抗体沉淀各组织PI4-K活力和酶蛋白的百分率基本相同,支持文献报道组织中微粒体或细胞膜中的PIK主要是PI4-K,而免疫性不同的PI3-K含量极少的观点.Based on the purification of phosphatidyl inositol-4-kinase (PI4-K) from porcine liver microsome,the antiserum of this enzyme was prepared from rabbits.The IgG of antiserum was purified,an ELISA of PI4-K was established, and the PI4-K in various tissues were immunologically analyzed and quantified.The results showed that the PI4-K in Triton X-100 solubilized microsomal membrane (TXSM) of porcine kidney and lung,Triton X-100 solublized porcine liver cell membrane and human neutrophil membrane could be precipitated with antiserum or antibody IgG of porcine liver microsonial PI4-K, revealing that there were common immunological determinants among the PI4-K from different tissues,subcelluler apparatus and species.The activities or enzyme protien amounts of PI4-K in the TXSM of porcine liver, kidney,lung,stomach.intestine and colon were sirnilar to each other.The amount and activity of PI4-K in the neutrophil membrane of acute neutrophilic leukemia patients were both increased to 3.5 fold as compared with that of normal aduhs.Additionally, in spite of different tissues,the percentages of PI4-K activity and enzyme protien precipitited by antibody of PI4-K were almost the same,supporting the reported concept in literature that the main PIK in microsome or cell membrane of the above tissues was PI4-K, and the contents of immunologically different PI3-Kwere very low in the above tissues.
关 键 词:免疫分析 ELISA 白血病 磷脂酰肌醇 激酶 PI-4-K
分 类 号:R392.11[医药卫生—免疫学] R733.710.2[医药卫生—基础医学]
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