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首页> 外文期刊>Cell motility and the cytoskeleton >Linkage of a nucleolin-related protein and casein kinase II with the detergent-stable photoreceptor cytoskeleton.
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Linkage of a nucleolin-related protein and casein kinase II with the detergent-stable photoreceptor cytoskeleton.

机译：核仁素相关蛋白和酪蛋白激酶II与去污剂稳定的感光细胞骨架的联系。

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摘要

Vertebrate photoreceptors are highly polarized sensory neurons with a complex microtubule and actin-based cytoskeletal organization. In the present study, we have used a detergent-extracted cytokeleton preparation from bovine photoreceptors to test the hypothesis that protein kinases and their substrates co-purify with the photoreceptor cytoskeleton. We incubated the cytoskeletal preparation in the presence of [gamma-32P]ATP. Following SDS-PAGE and autoradiography, we found two principal phosphoproteins with apparent molecular weights of 55 kDa (pp55) and 112 kDa (pp112). We have additionally identified the kinase responsible for phosphorylation of pp112 (and possibly pp55) as a casein kinase II-like enzyme. pp55 was identified as beta-tubulin based on Western blotting and its position on two-dimensional gels. Microsequencing revealed that 16 of the first 17 amino acids of pp112 were identical to human nucleolin, a nuclear protein. Western blotting, mobility in SDS PAGE and in two-dimensional gels, predominant localization within the nucleus, and phosphorylation by a casein kinase II all support the conclusion that pp112 is a nucleolin-related protein. Immunocytochemistry revealed a significant extranuclear pool of nucleolin-immunoreactivity within the cell bodies of photoreceptors. These findings suggest an important extranuclear role for nucleolin or a related protein in photoreceptors.
机译：脊椎动物的感光细胞是高度极化的感觉神经元，具有复杂的微管和基于肌动蛋白的细胞骨架组织。在本研究中，我们使用了从牛感光细胞中提取去污剂的细胞骨架，以检验蛋白质激酶及其底物与感光细胞骨架共同纯化的假说。我们在[γ-32P] ATP存在下孵育了细胞骨架制剂。经过SDS-PAGE和放射自显影后，我们发现了两种主要的磷蛋白，其表观分子量分别为55 kDa（pp55）和112 kDa（pp112）。我们还确定了负责pp112（可能还有pp55）磷酸化的激酶为酪蛋白激酶II样酶。根据Western印迹及其在二维凝胶上的位置，pp55被鉴定为β-微管蛋白。微量测序表明pp112的前17个氨基酸中有16个与人核仁素（一种核蛋白）相同。 Western印迹，在SDS PAGE和二维凝胶中的迁移性，在细胞核中的主要定位以及酪蛋白激酶II的磷酸化都支持pp112是与核蛋白相关的蛋白质的结论。免疫细胞化学揭示了光感受器细胞体内核仁免疫反应性的显着核外池。这些发现表明核仁蛋白或相关蛋白在感光细胞中具有重要的核外作用。

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