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Localization of palmitoylated and activated G protein α‐subunit in Dictyostelium discoideum Dictyostelium discoideum

机译:Dictyostelium discoideum dictyostelium discoideum中棕榈酰基化和活化G蛋白α-亚基的定位

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Abstract Guanine nucleotide‐binding proteins (G proteins) act as molecular switches to regulate many fundamental cellular processes. The lipid modification, palmitoylation, can be considered as a key factor for proper G protein function and plasma membrane localization. In Dictyostelium discoidum , Gα2 is essential for the chemotactic response to cAMP in their developmental life cycle. However, the regulation of Gα2 with respect to palmitoylation, activation and Gβγ association is less clear. In this study, Gα2 is shown to be palmitoylated on Cys‐4 by [ 3 H]palmitate labeling. Loss of this palmitoylation site results in redistribution of Gα2 within the cell and poor D. discoideum development. Cellular re‐localization is also observed for activated Gα2. In the membrane fraction, Gα2‐wt (YFP) is highly enriched in a low‐density membrane fraction, which is palmitoylation‐dependent. Activated Gα2 monomer and heterotrimer are shifted to two different higher‐density fractions. These results broaden our understanding of how G protein localization and function are regulated inside the cells.
机译:摘要鸟嘌呤核苷酸结合蛋白(G蛋白)作为分子开关来调节许多基本细胞过程。棕榈酰基棕榈酰基化,可被认为是适当G蛋白质功能和质膜定位的关键因素。在Dictyostelium Discopoidum中,Gα2对于在其发育生命周期中对阵营的趋化性反应至关重要。然而,关于棕榈酰基,活化和Gβγ关联的Gα2的调节较小。在该研究中,Gα2显示在Cys-4上是棕榈酰基化的[3 h]棕榈酸盐标记。该棕榈酰基化位点的丧失导致细胞内Gα2的再分布,差D. discoideum发育。对于活化的Gα2,还观察到细胞再定位。在膜级分中,Gα2-WT(YFP)高度富集在低密度膜馏分中,这是棕榈酰基依赖性的。活化的Gα2单体和异络剂被移至两种不同的高密度级分。这些结果拓宽了我们对细胞内部监管G蛋白定位和功能的理解。

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