PINK1 autophosphorylation is required for ubiquitin recognition
PINK1 autophosphorylation is required for ubiquitin recognition
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DOI：
10.15252/embr.201744981
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发表时间：
2018-04-01
期刊：
影响因子：
7.7
通讯作者：
Trempe, Jean-Francois
中科院分区：
文献类型：
--
作者：
Rasool, Shafqat;Soya, Naoto;Trempe, Jean-Francois
PINK 1突变导致常染色体隐性帕金森病（PD），一种神经退行性运动障碍。PINK 1是一种激酶，作为线粒体损伤的传感器，并启动受损细胞器的Parkin介导的清除。PINK 1磷酸化Parkin的泛素和泛素样（Ubl）结构域中的Ser 65，这刺激了其E3连接酶活性。PINK 1的自磷酸化是帕金激活所必需的，但这如何调节泛素激酶活性尚不清楚。在这里，我们表明赤拟谷盗PINK 1的自磷酸化是底物识别所必需的。使用酶动力学和NMR光谱，我们揭示了PINK 1通过一个保守的界面与帕金Ubl结合，其亲和力比泛素高10倍，该界面也与RING 1和SH 3结合有关。这种相互作用需要在Ser 205处磷酸化，Ser 205是一个不变的PINK 1残基（人类中为Ser 228）。使用质谱，我们证明了PINK 1在Ser 205处快速自磷酸化。小角X射线散射和氢-氘交换实验提供了对PINK 1催化结构域的深入了解。我们的研究结果表明，多个PINK 1分子在结合和磷酸化泛素和帕金蛋白之前首先自磷酸化。
Mutations in PINK1 cause autosomal recessive Parkinson's disease (PD), a neurodegenerative movement disorder. PINK1 is a kinase that acts as a sensor of mitochondrial damage and initiates Parkin-mediated clearance of the damaged organelle. PINK1 phosphorylates Ser65 in both ubiquitin and the ubiquitin-like (Ubl) domain of Parkin, which stimulates its E3 ligase activity. Autophosphorylation of PINK1 is required for Parkin activation, but how this modulates the ubiquitin kinase activity is unclear. Here, we show that autophosphorylation of Tribolium castaneum PINK1 is required for substrate recognition. Using enzyme kinetics and NMR spectroscopy, we reveal that PINK1 binds the Parkin Ubl with a 10-fold higher affinity than ubiquitin via a conserved interface that is also implicated in RING1 and SH3 binding. The interaction requires phosphorylation at Ser205, an invariant PINK1 residue (Ser228 in human). Using mass spectrometry, we demonstrate that PINK1 rapidly autophosphorylates in trans at Ser205. Small-angle X-ray scattering and hydrogen-deuterium exchange experiments provide insights into the structure of the PINK1 catalytic domain. Our findings suggest that multiple PINK1 molecules autophosphorylate first prior to binding and phosphorylating ubiquitin and Parkin.