Gosti F, Marty M C, Courvalin J C, Maunoury R, Bornens M
Proc Natl Acad Sci U S A. 1987 Feb;84(4):1000-4. doi: 10.1073/pnas.84.4.1000.
A spontaneously arising rabbit anti-centrosome serum with strong human specificity, used to identify specific antigens in isolated centrosomes, was shown to react with several noncentrosomal proteins including a 36-kDa protein that appeared to be the major cellular antigen. To explore the immunological relationship between noncentrosomal and centrosomal antigens, immunoglobulins were affinity purified using the individual noncentrosomal antigens (from lymphoblastoma KE37 cells) and were tested for their capacity to bind to human centrosomes in situ and to proteins from isolated centrosomes. In this way, the 36-kDa antigen, an abundant cytosolic protein, was shown to share at least one antigenic determinant with high molecular weight centrosomal proteins. This antigen was further identified by mild proteolysis as the glycolytic enzyme lactate dehydrogenase. In all the analyzed human cell lines, the centrosomal staining in situ was correlated with a strong labeling of purified lactate dehydrogenase in immunoblots. Conversely, the absence of centrosomal staining in rodent cells was always correlated with the absence of lactate dehydrogenase labeling. These data suggest an evolutionary relationship between centrosomal proteins and this "housekeeping" enzyme.
一种具有强人类特异性的自发产生的兔抗中心体血清，用于鉴定分离的中心体中的特定抗原，结果显示它能与几种非中心体蛋白发生反应，包括一种36 kDa的蛋白，该蛋白似乎是主要的细胞抗原。为了探究非中心体抗原与中心体抗原之间的免疫关系，使用单个非中心体抗原（来自淋巴瘤KE37细胞）对免疫球蛋白进行亲和纯化，并检测它们原位结合人中心体以及与分离的中心体蛋白结合的能力。通过这种方式，发现这种36 kDa的抗原，一种丰富的胞质蛋白，与高分子量的中心体蛋白至少共享一个抗原决定簇。通过温和的蛋白酶解进一步鉴定该抗原为糖酵解酶乳酸脱氢酶。在所有分析的人类细胞系中，原位中心体染色与免疫印迹中纯化的乳酸脱氢酶的强标记相关。相反，啮齿动物细胞中没有中心体染色总是与没有乳酸脱氢酶标记相关。这些数据表明中心体蛋白与这种“管家”酶之间存在进化关系。
