Functional properties of LRRK2 mutations in Taiwanese Parkinson disease

Kuo Hsuan Chang, Chiung Mei Chen, Chih Hsin Lin, Wen Teng Chang, Pei Ru Jiang, Ya Chin Hsiao, Yih Ru Wu, Guey Jen Lee-Chen

Research output: Contribution to journalArticle

Abstract

Background/purpose Leucine-rich repeat kinase 2 (LRRK2) is a large protein encoding multiple functional domains. Mutations within different LRRK2 domains have been considered to be involved in the development of Parkinson disease by different mechanisms. Our previous study found three LRRK2 mutations—p.R767H, p.S885N, and p.R1441H—in Taiwanese patients with Parkinson disease. Methods We evaluated the functional properties of LRRK2 p.R767H, p.S885N, and p.R1441H mutations by overexpressing them in human embryonic kidney 293 and neuroblastoma SK-N-SH cells. The common p.G2019S mutation in the kinase domain was included for comparison. Results In 293 cells, overexpressed p.R1441H—but not p.R767H, p.S885N, or p.G2019—increased GTP binding affinity to prolong the active state. Overexpressed p.R1441H and p.G2019S generated inclusions in 293 cells. In SK-N-SH cells, the α-synuclein was coexpressed with wild type as well as mutated p.R767H, p.S885N, p.R1441H, and p.G2019 LRRK2 proteins. Part of the perinuclear inclusions formed by p.R1441H and p.G2019S were colocalized with α-synuclein. Additionally, p.S885N and p.R1441H mutations caused reduced interaction between LRRK2 and ARHGEF7, a putative guanine nucleotide exchange factor for LRRK2, whereas this interaction was well preserved in p.R767H and p.G2019S mutations. Conclusion Our study suggests that p.R1441H protein facilitates the formation of intracellular inclusions, compromises GTP hydrolysis by increasing its affinity for GTP, and reduces its interaction with ARHGEF7.

Original languageEnglish
Pages (from-to)197-204
Number of pages8
JournalJournal of the Formosan Medical Association
Volume116
Issue number3
DOIs
Publication statusPublished - 2017 Mar 1

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Leucine
Parkinson Disease
Phosphotransferases
Mutation
Synucleins
Guanosine Triphosphate
Guanine Nucleotide Exchange Factors
Neuroblastoma
Protein Kinases
Proteins
Hydrolysis
Kidney

Keywords

  • ARHGEF7
  • GTPase
  • LRRK2
  • p.R1441H mutation
  • Parkinson disease

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Functional properties of LRRK2 mutations in Taiwanese Parkinson disease. / Chang, Kuo Hsuan; Chen, Chiung Mei; Lin, Chih Hsin; Chang, Wen Teng; Jiang, Pei Ru; Hsiao, Ya Chin; Wu, Yih Ru; Lee-Chen, Guey Jen.

In: Journal of the Formosan Medical Association, Vol. 116, No. 3, 01.03.2017, p. 197-204.

Research output: Contribution to journalArticle

Chang, Kuo Hsuan ; Chen, Chiung Mei ; Lin, Chih Hsin ; Chang, Wen Teng ; Jiang, Pei Ru ; Hsiao, Ya Chin ; Wu, Yih Ru ; Lee-Chen, Guey Jen. / Functional properties of LRRK2 mutations in Taiwanese Parkinson disease. In: Journal of the Formosan Medical Association. 2017 ; Vol. 116, No. 3. pp. 197-204.
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AU - Jiang, Pei Ru

AU - Hsiao, Ya Chin

AU - Wu, Yih Ru

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AB - Background/purpose Leucine-rich repeat kinase 2 (LRRK2) is a large protein encoding multiple functional domains. Mutations within different LRRK2 domains have been considered to be involved in the development of Parkinson disease by different mechanisms. Our previous study found three LRRK2 mutations—p.R767H, p.S885N, and p.R1441H—in Taiwanese patients with Parkinson disease. Methods We evaluated the functional properties of LRRK2 p.R767H, p.S885N, and p.R1441H mutations by overexpressing them in human embryonic kidney 293 and neuroblastoma SK-N-SH cells. The common p.G2019S mutation in the kinase domain was included for comparison. Results In 293 cells, overexpressed p.R1441H—but not p.R767H, p.S885N, or p.G2019—increased GTP binding affinity to prolong the active state. Overexpressed p.R1441H and p.G2019S generated inclusions in 293 cells. In SK-N-SH cells, the α-synuclein was coexpressed with wild type as well as mutated p.R767H, p.S885N, p.R1441H, and p.G2019 LRRK2 proteins. Part of the perinuclear inclusions formed by p.R1441H and p.G2019S were colocalized with α-synuclein. Additionally, p.S885N and p.R1441H mutations caused reduced interaction between LRRK2 and ARHGEF7, a putative guanine nucleotide exchange factor for LRRK2, whereas this interaction was well preserved in p.R767H and p.G2019S mutations. Conclusion Our study suggests that p.R1441H protein facilitates the formation of intracellular inclusions, compromises GTP hydrolysis by increasing its affinity for GTP, and reduces its interaction with ARHGEF7.

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