GTP binding is essential to the protein kinase activity of LRRK2, a causative gene product for familial Parkinson's disease
GTP binding is essential to the protein kinase activity of LRRK2, a causative gene product for familial Parkinson's disease
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DOI：
10.1021/bi061960m
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发表时间：
2007-02-06
期刊：
影响因子：
2.9
通讯作者：
Iwatsubo, Takeshi
中科院分区：
文献类型：
--
作者：
Ito, Genta;Okai, Takuro;Iwatsubo, Takeshi
富含亮氨酸重复激酶2（LRRK2）是家族性帕金森病常染色体显性遗传形式（PARK8）致病基因的产物，除激酶域外，它还含有一个类Ras小GTP结合蛋白结构域（ROC），不过这两个功能结构域之间的关系仍不明确。在此，我们通过薄层色谱分析表明，LRRK2能稳定结合GTP，但在HEK293和Neuro - 2a细胞中缺乏GTP酶活性。一种将LRRK2转变为无鸟嘌呤核苷酸形式的ROC结构域突变（T1348N），会消除LRRK2的激酶活性及其在代谢标记时的磷酸掺入。LRRK2的磷酸化受到环磷酸腺苷（cAMP）依赖性蛋白激酶潜在抑制剂的抑制。这些数据表明，GTP与ROC结构域的结合，既调控LRRK2的激酶活性，也调控其被其他激酶磷酸化的过程。
Leucine-rich repeat kinase 2 (LRRK2), a product of a causative gene for the autosomal-dominant form of familial Parkinson's disease (PARK8), harbors a Ras-like small GTP binding protein-like (ROC) domain besides the kinase domain, although the relationship between these two functional domains remains elusive. Here we show by thin-layer chromatographic analysis that LRRK2 stably binds GTP but lacks a GTPase activity in HEK293 and Neuro-2a cells. A ROC domain mutation that converts LRRK2 to a guanine nucleotide-free form (T1348N) abolishes the kinase activity of LRRK2 as well as its phosphate incorporation upon metabolic labeling. The phosphorylation of LRRK2 was inhibited by potential inhibitors for cyclic AMP-dependent protein kinase. These data suggest that binding of GTP to the ROC domain regulates the kinase activity of LRRK2 as well as its phosphorylation by other kinase(s).