Antisera inhibiting mammalian cell spreading and possible cell surface antigens involved.
Antisera inhibiting mammalian cell spreading and possible cell surface antigens involved.
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DOI：
10.1083/jcb.86.3.866
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发表时间：
1980-09
期刊：
影响因子：
--
通讯作者：
Sueoka N
中科院分区：
文献类型：
--
作者：
Hsieh P;Sueoka N
用活细胞静脉注射法制备了抗大鼠神经元肿瘤细胞系(B103)的抗血清。这种免疫血清(抗B103)在15种不同的大鼠神经细胞系以及小鼠和大鼠成纤维细胞系中沉淀出一些细胞表面蛋白，经双向凝胶电泳法可识别为共同的放射性碘化斑点。用十二烷基硫酸钠凝胶电泳法估计，一种主要常见蛋白质(II4)的表观分子量在80,000到90,000之间，另一种蛋白质(I3)的表观分子量为120,000。这两种蛋白在不同的细胞系中都能被该抗血清识别。此外，在1：20的稀释度下，该血清可使单层细胞在0.5h内聚集，并在3h内从平板上分离。它还能抑制新培养细胞的扩散。抗血清的这些作用是可逆的。当抗血清被细胞吸收(例如，被胶质细胞系B27吸收)时，该血清不再引起单层细胞的聚集，它不抑制细胞扩散，它不从各种细胞系的细胞表面免疫沉淀这两种共同的蛋白质。针对其他几个大鼠细胞系的抗血清也从细胞表面沉淀出相同的共同蛋白(II4和I3)，并防止细胞扩散。这些数据表明，抗体首先作用于细胞表面，然后抑制细胞扩散或聚集扩散的细胞。免疫沉淀的细胞表面蛋白在双向凝胶电泳图上的一致模式使这两个共同的表面蛋白(II4或I3或两者都有)可能成为抗体结合的靶蛋白。因此，它们可能在细胞扩散中发挥关键作用。
Antiserum against a rat neuronal tumor cell line (B103) has been prepared in rabbit by intravenous injection of live cells. This immune serum (anti-B103) precipitates a few cell surface proteins recognizable by two-dimensional gel electrophoresis as common radioiodinatable spots in 15 different rat neural cell lines and in mouse and rat fibroblast cell lines. The apparent molecular weight of one major common protein (II4) is estimated by SDS gel electrophoresis to be somewhere between 80,000 and 90,000 and another protein (I3) to be 120,000. These two proteins are consistently recognized in various cell lines by this antiserum. Furthermore, at a 1:20 dilution, this serum causes monolayer cells to round up usually within 0.5 h and detach from the plate within 3 h. It also inhibits spreading of freshly plated cells. These effects of the antiserum are reversible. Upon absorption of the antiserum with cells (e.g., absorbed with a glial cell line, B27), the serum no longer causes the rounding up of the monolayer cells, it does not inhibit cell spreading, and it does not immune-precipitate the two common proteins from the cell surface of various cell lines. Antisera against several other rat cell lines also precipitate the same common proteins (II4 and I3) from the cell surface and prevent cell spreading. These data suggest that the antibody acts first at the cell surface and then inhibits cell spreading or rounding up of spread cells. The consistent pattern of the immunoprecipitated cell surface proteins on the two- dimensional gel electrophoresis makes these two common surface proteins (II4 or I3 or both) possible candidates for target proteins to which the antibody binds. Thus, they may play a critical role in cell spreading.