Lampidis T J, Trevorrow K W, Rubin R W
Exp Cell Res. 1986 Jun;164(2):463-70. doi: 10.1016/0014-4827(86)90044-3.
The effects of the tubulin-binding drug colchicine on cultured neonate cardiac cell function were investigated. Application of low doses of colchicine (but not lumicolchicine) caused an early reversible increase in beating rate with a concomitant decrease in amplitude. Treatment of the cells with trypsin at a dose that removes surface tubulin but does not inhibit spontaneous beating, diminished the colchicine effect. Surface radio-iodination of the live cultures followed by two-dimensional gel electrophoresis and radioautography revealed that two spots were heavily labeled. These spots co-migrated with purified brain tubulin. Fibroblasts derived from the cardiac cultures did not label over the tubulin spots. Trypsin treatment removed the presumptive tubulin from the radioautographs but only removed the most basic portion of the alpha-tubulin spot from the stained gel. These results are consistent with a surface membrane role for an iso-form of tubulin in neonate cardiac cells.
研究了微管蛋白结合药物秋水仙碱对培养的新生心肌细胞功能的影响。应用低剂量的秋水仙碱(而非光秋水仙碱)会导致早期可逆的心率增加,同时振幅降低。用能去除表面微管蛋白但不抑制自发搏动的剂量的胰蛋白酶处理细胞,会减弱秋水仙碱的作用。对活细胞培养物进行表面放射性碘化,然后进行二维凝胶电泳和放射自显影,结果显示有两个斑点被大量标记。这些斑点与纯化的脑微管蛋白共同迁移。源自心肌培养物的成纤维细胞在微管蛋白斑点处未被标记。胰蛋白酶处理从放射自显影片中去除了假定的微管蛋白,但仅从染色凝胶中去除了α-微管蛋白斑点最碱性的部分。这些结果与微管蛋白同工型在新生心肌细胞表面膜中的作用一致。