Greggio Elisa, Taymans Jean-Marc, Zhen Eugene Yuejun, Ryder John, Vancraenenbroeck Renée, Beilina Alexandra, Sun Peng, Deng Junpeng, Jaffe Howard, Baekelandt Veerle, Merchant Kalpana, Cookson Mark R
Cell Biology and Gene Expression Unit, Laboratory of Neurogenetics, National Institute on Aging, Bethesda, MD 20892-3707, USA.
Biochem Biophys Res Commun. 2009 Nov 20;389(3):449-54. doi: 10.1016/j.bbrc.2009.08.163. Epub 2009 Sep 3.
Mutations in leucine-rich repeat kinase 2 (LRRK2) are a common cause of inherited Parkinson's disease (PD). The protein is large and complex, but pathogenic mutations cluster in a region containing GTPase and kinase domains. LRRK2 can autophosphorylate in vitro within a dimer pair, although the significance of this reaction is unclear. Here, we mapped the sites of autophosphorylation within LRRK2 and found several potential phosphorylation sites within the GTPase domain. Using mass spectrometry, we found that Thr1343 is phosphorylated and, using kinase dead versions of LRRK2, show that this is an autophosphorylation site. However, we also find evidence for additional sites in the GTPase domain and in other regions of the protein suggesting that there may be multiple autophosphorylation sites within LRRK2. These data suggest that the kinase and GTPase activities of LRRK2 may exhibit complex autoregulatory interdependence.
富含亮氨酸重复激酶2（LRRK2）的突变是遗传性帕金森病（PD）的常见病因。该蛋白体积大且结构复杂，但致病突变集中在包含GTP酶和激酶结构域的区域。LRRK2可在体外二聚体对中进行自磷酸化，尽管该反应的意义尚不清楚。在这里，我们绘制了LRRK2内自磷酸化的位点，并在GTP酶结构域内发现了几个潜在的磷酸化位点。使用质谱分析，我们发现苏氨酸1343被磷酸化，并且使用LRRK2的激酶失活版本，表明这是一个自磷酸化位点。然而，我们也在GTP酶结构域和该蛋白的其他区域发现了其他位点的证据，这表明LRRK2内可能存在多个自磷酸化位点。这些数据表明，LRRK2的激酶和GTP酶活性可能表现出复杂的自调节相互依赖性。
