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一种大鼠肝脏57千道尔顿蛋白被鉴定与他汀类药物(一种非增殖细胞的标志物)共享抗原决定簇。

A rat liver 57-kDa protein is identified to share antigenic determinants with statin, a marker for nonproliferating cells.

作者信息

Sester U, Moutsatsos I K, Wang E

机构信息

Bloomfield Centre for Research in Aging, Sir Mortimer B. Davis-Jewish General Hospital, Montreal, Quebec, Canada.

出版信息

Exp Cell Res. 1989 Jun;182(2):550-8. doi: 10.1016/0014-4827(89)90258-9.

Abstract

The nuclear protein statin, found uniquely in nonproliferating cells, has been previously identified by mouse monoclonal antibodies designated as S-30 and S-44. We report here the screening of various rat tissues for proteins cross-reacting with these antibodies. As revealed by immunoblotting, two polypeptides (mol wt 80 and 57 kDa) and a group of lower-molecular-weight proteins migrating between 34 and 38 kDa were found to react with the anti-statin antibodies. The most prominent immunoreactivity was observed with a 57-kDa protein present in rat liver. Upon further fractionation of the liver protein extract with ammonium sulfate [(NH4)2SO4] the 57-kDa protein, designated as rat liver protein 57 or RLp57, was detected independently with both anti-statin antibodies in the 30 to 60% (NH4)2SO4 fraction. In order to determine whether rat liver protein 57 is indeed specifically recognized by anti-statin antibody S-44, we used RLp57 transferred onto nitrocellulose paper as a specific substrate for the adsorption of the S-44 antibody from ascites fluid. As shown by indirect immunofluorescence microscopy the solution remaining after adsorption failed to stain human fibroblasts. The adsorbed immunoglobulin, however, upon elution revealed statin-specific nuclear staining activity on senescent fibroblasts. These findings suggest that rat liver protein 57 and the human fibroblast statin share similar antigenic determinants recognized by the statin-specific S-44 antibody. Our results indicate furthermore that the statin previously identified in fibroblasts may represent one member of a group of several antigenically related proteins detectable with specific anti-statin antibodies.

摘要

核蛋白他汀仅在非增殖细胞中发现,此前已通过名为S - 30和S - 44的小鼠单克隆抗体鉴定出来。我们在此报告了对各种大鼠组织进行筛选,以寻找与这些抗体发生交叉反应的蛋白质。免疫印迹显示,发现两种多肽(分子量分别为80 kDa和57 kDa)以及一组分子量在34至38 kDa之间迁移的低分子量蛋白质与抗他汀抗体发生反应。在大鼠肝脏中存在的一种57 kDa蛋白质观察到最显著的免疫反应性。用硫酸铵[(NH4)2SO4]对肝脏蛋白提取物进行进一步分级分离后,在30%至60%(NH4)2SO4级分中,两种抗他汀抗体均独立检测到了这种被命名为大鼠肝脏蛋白57或RLp57的57 kDa蛋白质。为了确定大鼠肝脏蛋白57是否确实被抗他汀抗体S - 44特异性识别,我们将RLp57转移到硝酸纤维素纸上,作为从腹水液中吸附S - 44抗体的特异性底物。间接免疫荧光显微镜检查显示,吸附后剩余的溶液未能对人成纤维细胞进行染色。然而,洗脱后的吸附免疫球蛋白在衰老的成纤维细胞上显示出他汀特异性的核染色活性。这些发现表明,大鼠肝脏蛋白57和人成纤维细胞他汀具有相似的抗原决定簇,可被他汀特异性S - 44抗体识别。我们的结果还表明,先前在成纤维细胞中鉴定出的他汀可能代表了一组可被特异性抗他汀抗体检测到的、具有抗原相关性的蛋白质中的一个成员。

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