Association of kinesin and myosin with pigment granules in crustacean chromatophores
Association of kinesin and myosin with pigment granules in crustacean chromatophores
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DOI：
10.1111/j.1600-0749.2005.00277.x
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发表时间：
2006-02-01
期刊：
影响因子：
--
通讯作者：
McNamara, JC
中科院分区：
文献类型：
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作者：
Boyle, RT;McNamara, JC
甲壳类动物的色彩适应是由神经分泌肽引起的细胞信号传导导致的色素细胞内有色色素颗粒的差异性分布引起的。然而，力转导，机械化学蛋白马达负责颗粒易位，和他们的分子作用机制，没有得到很好的理解。本研究采用免疫细胞化学技术和运动试验在体外证明，从驱动蛋白和肌球蛋白超家族的蛋白马达是稳定相关的红色，卵巢色素细胞的淡水，palaemonid虾，Macrobrachium olfersii的膜结合的色素颗粒。单克隆抗体对传统的驱动蛋白重链，和抗肌球蛋白的全血清，标记的色素含有片段的色素细胞的匀浆与完全分散或聚集的色素：这一发现推断蛋白马达和色素颗粒之间的永久协会，并表明，这种电机可能受到监管，而绑定到他们的货物。色素聚集体似乎是一种肌球蛋白，因为抗肌球蛋白全血清在体外运动试验中减弱了肌肉触发的色素聚集，并诱导了某些色素细胞中的色素过度分散。蛋白质印迹的色素，卵巢组织匀浆证明蛋白质带一致的肌球蛋白II和肌球蛋白XII，其中任何一个可能是色素聚集。这项研究提供了第一个直接的证据，肌球蛋白和驱动蛋白马达直接和稳定地与甲壳动物色素颗粒，并可能代表第一次成功分离肌球蛋白类XII。
Chromatic adaptation in crustaceans results from the differential distribution of colored pigment granules within their chromatophores consequent to cell signaling by neurosecretory peptides. However, the force transducing, mechanochemical protein motors responsible for granule translocation, and their molecular mechanisms of action, are not well understood. The present study uses immunocytochemical techniques and a motility assay in vitro to demonstrate that protein motors from the kinesin and myosin superfamilies are stably associated with membrane-bounded pigment granules in the red, ovarian chromatophores of the freshwater, palaemonid shrimp, Macrobrachium olfersii. Monoclonal antibodies against conventional kinesin heavy chain, and an anti-myosin whole serum, labeled pigment-containing fragments prepared from homogenates of chromatophores with fully dispersed or aggregated pigments: this finding infers a permanent association between the protein motors and the pigment granules, and suggests that such motors may be regulated while bound to their cargos. The pigment aggregator appears to be a myosin since the anti-myosin whole serum attenuated hormonally triggered pigment aggregation in the motility assay in vitro, and induced pigment hyper-dispersion in some chromatophores. Western blots of the chromatophore-containing, ovarian tissue homogenate demonstrated protein bands consistent with myosin II and myosin XII, either of which may be the pigment aggregator. This study provides the first direct evidence for myosin and kinesin protein motors directly and stably associated with pigment granules in crustacean chromatophores, and may represent the first successful isolation of myosin class XII.