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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >FTDP-17 missense mutations site-specifically inhibit as well as promote dephosphorylation of microtubule-associated protein tau by protein phosphatases of HEK-293 cell extract.
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FTDP-17 missense mutations site-specifically inhibit as well as promote dephosphorylation of microtubule-associated protein tau by protein phosphatases of HEK-293 cell extract.

机译:FTDP-17错义突变位点特异性抑制HEK-293细胞提取物的蛋白磷酸酶,并促进微管相关蛋白tau的去磷酸化。

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摘要

FTDP-17 missense tau mutations: G272V, P301L, V337M and R406W promote tau phosphorylation in human and transgenic mice brains by interfering with the tau phosphorylation/dephosphorylation balance. The effect of FTDP-17 mutations on tau phosphorylation by different kinases has been studied previously. However, it is not known how various FTDP-17 mutations affect tau dephosphorylation by phosphoprotein phosphatases. In this study we have observed that when transfected into HEK-293 cells, tau is phosphorylated on various sites that are also phosphorylated in diseased human brains. When transfected cells are lysed and incubated, endogenously phosphorylated tau is dephosphorylated by cellular protein phosphatase 1 (PP1), phosphatase 2A (PP2A) and phosphatase 2B (PP2B), which are also present in the lysate. By using this assay and specific inhibitors of PP1, PP2A and PP2B, we have observed that the G272V mutation promotes tau dephosphorylation by PP2A at Ser(396/404), Ser(235), Thr(231), Ser(202/205) and Ser(214) and by PP2B at Ser(214) but inhibits dephosphorylation by PP2B at Ser(396/404). The P301L mutation promotes tau dephosphorylation at Thr(231) by PP1 and at Ser(396/404), Thr(231), Ser(235) and Ser(202/205) by PP2A but inhibits dephosphorylation at Ser(214) by PP2B. The V337M mutation promotes tau dephosphorylation at Ser(235), Thr(231) and Ser(202/205) by PP2A and at Ser(202/205) by PP2B whereas the R406W mutation promotes tau dephosphorylation at Ser(396/404) by PP1, PP2A and PP2B but inhibits dephosphorylation at Ser(202/205) and Ser(235) by PP1 and PP2A, respectively. Our results indicate that each FTDP-17 tau mutation not only site-specifically inhibits tau dephosphorylation on some sites but also promotes dephosphorylation by phosphatases on other sites.
机译:FTDP-17错义tau突变:G272V,P301L,V337M和R406W通过干扰tau磷酸化/去磷酸化平衡来促进人和转基因小鼠大脑中的tau磷酸化。先前已经研究了FTDP-17突变对不同激酶对tau磷酸化的影响。但是,尚不清楚各种FTDP-17突变如何影响磷蛋白磷酸酶对tau的去磷酸化作用。在这项研究中,我们已经观察到,当将tau基因转染到HEK-293细胞中时,其在各种部位的磷酸化作用在患病的人脑中也被磷酸化。将转染的细胞裂解并温育后,内源磷酸化的tau会被细胞蛋白磷酸酶1(PP1),磷酸酶2A(PP2A)和磷酸酶2B(PP2B)磷酸化,后者也存在于裂解物中。通过使用该测定法和PP1,PP2A和PP2B的特异性抑制剂,我们观察到G272V突变可促进PP2A在Ser(396/404),Ser(235),Thr(231),Ser(202/205)处的tau脱磷酸化和Ser(214)以及在Ser(214)处被PP2B抑制,但在Ser(396/404)处被PP2B抑制去磷酸化。 P301L突变促进PP1在Thr(231)和Ser(396/404),Thr(231),Ser(235)和Ser(202/205)处的tau脱磷酸作用,而由PP2B抑制Ser(214)的tau脱磷酸作用。 V337M突变通过PP2A促进Ser(235),Thr(231)和Ser(202/205)的tau去磷酸化,而PP2B在Ser(202/205)促进tau的去磷酸化,而R406W突变则通过PP2A促进Ser(396/404)的tau去磷酸化。 PP1,PP2A和PP2B,但分别抑制PP1和PP2A在Ser(202/205)和Ser(235)处的去磷酸化。我们的结果表明,每个FTDP-17 tau突变不仅在某些位点特异性抑制tau的去磷酸化,而且在其他位点上通过磷酸酶促进去磷酸化。

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