磷脂酰肌醇－4－激酶的的免疫分析

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．

Immunological Analysis of Phosphatidyl Inositol-4-kinase

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.