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首页> 外文期刊>Molecular and Cellular Biology >The Phosducin-Like Protein PhLP1 Is Essential for Gβγ Dimer Formation in Dictyostelium discoideum
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The Phosducin-Like Protein PhLP1 Is Essential for Gβγ Dimer Formation in Dictyostelium discoideum

机译：类视盘蛋白类蛋白PhLP1是盘基网柄菌中Gβγ二聚体形成所必需的。

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Phosducin proteins are known to inhibit G protein-mediated signaling by sequestering Gβγ subunits. However, Dictyostelium discoideum cells lacking the phosducin-like protein PhLP1 display defective rather than enhanced G protein signaling. Here we show that green fluorescent protein (GFP)-tagged Gβ (GFP-Gβ) and GFP-Gγ subunits exhibit drastically reduced steady-state levels and are absent from the plasma membrane in phlp1? cells. Triton X-114 partitioning suggests that lipid attachment to GFP-Gγ occurs in wild-type cells but not in phlp1? and gβ? cells. Moreover, Gβγ dimers could not be detected in vitro in coimmunoprecipitation assays with phlp1? cell lysates. Accordingly, in vivo diffusion measurements using fluorescence correlation spectroscopy showed that while GFP-Gγ proteins are present in a complex in wild-type cells, they are free in phlp1? and gβ? cells. Collectively, our data strongly suggest the absence of Gβγ dimer formation in Dictyostelium cells lacking PhLP1. We propose that PhLP1 serves as a cochaperone assisting the assembly of Gβ and Gγ into a functional Gβγ complex. Thus, phosducin family proteins may fulfill hitherto unsuspected biosynthetic functions.
机译：已知膦丝蛋白通过隔离Gβγ亚基来抑制G蛋白介导的信号传导。但是，缺乏类光导蛋白蛋白PhLP1的细胞表现出缺陷，而不是增强的G蛋白信号传导。在这里，我们显示带有绿色荧光蛋白（GFP）标签的Gβ（GFP-Gβ）和GFP-Gγ亚基表现出大大降低的稳态水平，并且在 phlp1 的质膜中不存在？ 单元格。 Triton X-114分区表明脂质附着到GFP-Gγ发生在野生型细胞中，而不发生在 phlp1 ？和gβ 中？单元格。此外，在 phlp1 ？细胞裂解液的共免疫沉淀试验中，无法在体外检测到Gβγ二聚体。因此，使用荧光相关光谱的体内扩散测量表明，虽然GFP-Gγ蛋白存在于野生型细胞的复合物中，但它们在 phlp1 ？和< em>gβ ？细胞。总体而言，我们的数据强烈表明缺乏PhLP1的细胞中不存在Gβγ二聚体。我们建议PhLP1充当辅助分子，协助将Gβ和Gγ组装成功能性Gβγ复合物。因此，光导蛋白家族蛋白可实现迄今未曾怀疑的生物合成功能。

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