Characterization of Rab phosphorylation by LRRK2 using Phos-tag SDS–PAGE
2019 Volume 63 Issue 2 Pages 31-34
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2019 Volume 63 Issue 2 Pages 31-34
LRRK2 (leucine-rich repeat kinase 2) is one of the causative gene products for familial Parkinson’s disease (PD), harboring a Ser/Thr kinase domain with unknown functions. Recently we identified several small GTPase Rab proteins including Rab10 as physiological substrates of LRRK2, and we found that LRRK2 mutations linked with PD abnormally increase the Rab10 phosphorylation in cells and tissues. We utilized Phos-tag SDS–PAGE for detecting the phosphorylation of endogenous Rab10. On Phos-tag SDS–PAGE gels, one can detect protein phosphorylation as a bandshift, which enabled us to detect the endogenous levels of Rab10 phosphorylation without generating phospho-specific antibodies. In this review, we summarized the recent Phos-tag analysis on Rab10 phosphorylation by LRRK2.
