Ogawa K, Hosoya H, Yokota E, Kobayashi T, Wakamatsu Y, Ozato K, Negishi S, Obika M
Eur J Cell Biol. 1987 Feb;43(1):3-9.
Platyfish-swordtail hybrid melanoma cells exhibit pigment aggregation in response to adrenergic stimulation or melanophore-concentrating hormone. This translocation of pigment granules is thought to be related to radially arrayed microtubules. Very little is known about the molecular "motor" that powers the translocation. We present evidence that dynein is located on these microtubules and is a candidate for the "motor". Vanadate and erythro-9-[3-(2-hydroxynonyl)]adenine, which are potent inhibitors of dynein ATPase, prevent the transport of melanosome granules in Brij-treated melanoma cells. Direct identification of dynein in melanoma cells and tissues is demonstrated by immunofluorescence microscopy and immunoblotting using anti-fragment A (tryptic fragment of sea urchin sperm dynein) serum. The cytoplasm of melanoma cells is stained with the antiserum and gives rise to a pattern similar to the distribution of microtubules. Western blotting shows that the molecular weight of an immunoreactive polypeptide in melanoma tissues coincides with that of the heavy chain of sea urchin sperm dynein.
剑尾鱼 - 新月鱼杂交黑色素瘤细胞在肾上腺素能刺激或促黑素细胞聚集激素作用下会出现色素聚集。这种色素颗粒的易位被认为与呈放射状排列的微管有关。对于驱动这种易位的分子“马达”,人们了解甚少。我们提供的证据表明,动力蛋白位于这些微管上,并且是“马达”的候选者。钒酸盐和erythro - 9 - [3 - (2 - 羟基壬基)]腺嘌呤是动力蛋白ATP酶的有效抑制剂,它们可阻止经Brij处理的黑色素瘤细胞中黑素体颗粒的运输。通过免疫荧光显微镜检查和使用抗片段A(海胆精子动力蛋白的胰蛋白酶片段)血清进行免疫印迹,直接鉴定了黑色素瘤细胞和组织中的动力蛋白。黑色素瘤细胞的细胞质被抗血清染色,呈现出与微管分布相似的模式。蛋白质印迹显示,黑色素瘤组织中一种免疫反应性多肽的分子量与海胆精子动力蛋白重链的分子量一致。