Lack of correlation between the kinase activity of LRRK2 harboring kinase-modifying mutations and its phosphorylation at Ser910, 935, and Ser955

Genta Ito; Tetta Fujimoto; Shogo Kamikawaji; Tomoki Kuwahara; Takeshi Iwatsubo

doi:10.1371/journal.pone.0097988

Lack of correlation between the kinase activity of LRRK2 harboring kinase-modifying mutations and its phosphorylation at Ser910, 935, and Ser955

PLoS One. 2014 May 16;9(5):e97988. doi: 10.1371/journal.pone.0097988. eCollection 2014.

Authors

Genta Ito¹, Tetta Fujimoto¹, Shogo Kamikawaji², Tomoki Kuwahara¹, Takeshi Iwatsubo³

Affiliations

¹ Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
² Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
³ Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.

Abstract

Leucine-rich repeat kinase 2 (LRRK2) is extensively phosphorylated in cells within a region amino-terminal to the leucine-rich repeat domain. Since phosphorylation in this region of LRRK2, including Ser910, Ser935, Ser955, and Ser973, is significantly downregulated upon treatment with inhibitors of LRRK2, it has been hypothesized that signaling pathways downstream of the kinase activity of LRRK2 are involved in regulating the phosphorylation of LRRK2, although the precise mechanism has remained unknown. Here we examined the effects of LRRK2 inhibitors on the phosphorylation state at Ser910, Ser935, and Ser955 in a series of kinase-inactive mutants of LRRK2. We found that the responses of LRRK2 to the inhibitors varied among mutants, in a manner not consistent with the above-mentioned hypothesis. Notably, one of the kinase-inactive mutants, T2035A LRRK2, underwent phosphorylation, as well as the inhibitor-induced dephosphorylation, at Ser910, Ser935, and Ser955, to a similar extent to those observed with wild-type LRRK2. These results suggest that the kinase activity of LRRK2 is not involved in the common mechanism of inhibitor-induced dephosphorylation of LRRK2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
HEK293 Cells
Humans
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Molecular Sequence Data
Mutation, Missense*
Phosphorylation
Protein Kinase Inhibitors / pharmacology
Protein Processing, Post-Translational*
Protein Serine-Threonine Kinases / antagonists & inhibitors
Protein Serine-Threonine Kinases / chemistry
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism*
Protein Structure, Tertiary
Serine / genetics
Serine / metabolism
Signal Transduction*

Substances

Protein Kinase Inhibitors
Serine
LRRK2 protein, human
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Protein Serine-Threonine Kinases

Grants and funding

This work was supported by the Japan Society for the Promotion of Science [grant number 22790058 and 24790068 for GI] [Grant-in-Aid for Young Scientists (B)], by the Ministry of Education, Culture, Sports, Science and Technology [grant number 17025009 for TI] (Grant-in-Aid for Scientific Research on Priority Areas-Research on Pathomechanism of Brain Disorders, and Global Center of Excellence Program), by the Japan Science and Technology Agency [Core Research for Evolutional Science and Technology (CREST) for TI] and by Elan Pharmaceuticals (Innovation Program). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.