APPEARANCE OF CYTOSKELETAL COMPONENTS ON THE SURFACE OF LEUKEMIA-CELLS AND OF LYMPHOCYTES TRANSFORMED BY MITOGENS AND EPSTEIN-BARR VIRUS
APPEARANCE OF CYTOSKELETAL COMPONENTS ON THE SURFACE OF LEUKEMIA-CELLS AND OF LYMPHOCYTES TRANSFORMED BY MITOGENS AND EPSTEIN-BARR VIRUS
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DOI：
10.1073/pnas.77.8.4979
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
RAPAPORT, FT
中科院分区：
文献类型：
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作者：
BACHVAROFF, RJ;MILLER, F;RAPAPORT, FT
标记蛋白质的乳过氧化物酶碘标记和2-维电泳已经证明了良好的特征的细胞骨架蛋白（肌动蛋白和微管蛋白）的表面上的人淋巴细胞进行blastogenic转化和某些恶性人类细胞。这种蛋白质不能在正常的静息人类淋巴细胞表面检测到。有丝分裂原转化的T和B淋巴细胞表面膜上最突出的细胞骨架蛋白是肌动蛋白。在爱泼斯坦-巴尔病毒基因组阳性伯基特淋巴瘤和淋巴母细胞样细胞系以及2种白血病细胞中，主要的碘化膜蛋白组分是肌动蛋白以及α 1-、α 2-和β-微管蛋白这些蛋白质牢固地连接到细胞质骨架上，不能被Triton X-100去除。同时用特异性抗体和F（ab“）2片段进行的免疫荧光研究证实了活的和固定的淋巴细胞表面上的细胞骨架成分的出现，与生化数据平行，并表明这种细胞骨架蛋白在细胞表面上形成了独特的模式，从小的斑块到大的突起。用[35 S]甲硫氨酸标记5小时表明，所有这些细胞释放大量标记的肌动蛋白和微管蛋白到培养基中。这些材料没有再吸附到细胞的膜表面。
Lactoperoxidase iodination and 2-dimensional electrophoresis of the labeled proteins have demonstrated well-characterized cytoskeletal proteins (actin and tubulins) on the surface of human lymphocytes undergoing blastogenic transformation and of certain malignant human cells. Such proteins could not be detected on the surface of normal resting human lymphocytes. The most prominent cytoskeletal protein identified on the surface membrane of mitogen-transformed T and B lymphocytes was actin. In Epstein-Barr virus genome-positive Burkitt''s lymphoma and lymphoblastoid cell lines and in 2 leukemia cells, the major iodinated membrane protein components were actin and .alpha.1-, .alpha.2- and .beta.-tubulins. These proteins were firmly connected to the cytoplasmic skeleton and could not be removed by Triton X-100. Concurrent immunofluorescence studies with specific antibodies and F(ab'')2 fragments confirmed the appearance of cytoskeletal components on the surface of live and fixed lymphocytes, in parallel with the biochemical data, and indicated that such cytoskeletal proteins formed distinctive patterns on the cell surface, ranging from small patches to large projections. Five hour labeling with [35S]methionine indicates that all such cells released large quantities of labeled actin and tubulins into the culture medium. These materials were not readsorbed to the membrane surfaces of the cells.