Actin-binding specificity of marine macrolide toxins, mycalolide B and kabiramide D.
Actin-binding specificity of marine macrolide toxins, mycalolide B and kabiramide D.
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海洋大环内酯毒素、mycalolide B 和 kabiramide D 的肌动蛋白结合特异性。
DOI：
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发表时间：
1998
期刊：
影响因子：
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通讯作者：
S. Watabe
中科院分区：
文献类型：
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作者：
S. Wada;S. Matsunaga;S. Saito;N. Fusetani;S. Watabe
肌动蛋白解聚的海洋天然产物，mycalcohol B，和相关化合物，kabiramide D，标记与biocytin，生物素衍生物，并用于指定培养的大鼠3 Y1成纤维细胞中的靶分子。Mycalcium B表现出与各种细胞内蛋白质结合的能力，可能是通过在Mycalcium B的C5上Michael加成巯基。然而，没有细胞内的蛋白质以外的肌动蛋白明显与biocytinylated kabiramide D反应，表明kabiramide D的结合肌动蛋白是高度特异性的。用生物胞素化的kabiramide D处理细胞，然后用异硫氰酸荧光素缀合的抗生物素蛋白染色，结果表明，生物胞素化的kabiramide D与由F-肌动蛋白组成的应力纤维结合，尽管染色强度弱于荧光鬼笔环肽染色。卡比胺D与肌动蛋白结合的试验表明，卡比胺D与bistheonelide A的肌动蛋白结合位点相同，而与latrunculin A和细胞松弛素D的结合位点不同。
An actin-depolymerizing marine natural product, mycalolide B, and a related compound, kabiramide D, were labeled with biocytin, a biotin derivative, and used to specify target molecules in cultured rat 3Y1 fibroblasts. Mycalolide B exhibited the ability to bind to various intracellular proteins, probably through the Michael addition of a sulfhydryl group to C5 of mycalolide B. However, no intracellular proteins other than actin apparently reacted with biocytinylated kabiramide D, demonstrating that the binding of kabiramide D to actin was highly specific. Cells treated with biocytinylated kabiramide D followed by staining with fluorescein isothiocyanate-conjugated avidin showed that biocytinylated kabiramide D bound to stress fibers composed of F-actin, although the staining intensity was weaker than the fluorescent phalloidin staining. The assay for the binding of kabiramide D to actin, which had previously been treated with other actin-depolymerizing agents, showed that the actin-binding site for kabiramide D was the same as that for bistheonellide A, but not those for latrunculin A and cytochalasin D.