TITLE:
Lack of Evidence for Decreased Protein Stability in the 2397 (Met) Haplotype of the Leucine Rich Repeat Kinase 2 Protein Implicated in Parkinson’s Disease
AUTHORS:
Ryan S. Anderton, Lauren Hill, Rhiannon Morris, Frank L. Mastaglia, Wayne Greene, Sherif Boulos
KEYWORDS:
Parkinson’s Disease, LRRK2, Met2397, Thr2397, A1442P, R1441C
JOURNAL NAME:
Advances in Parkinson's Disease,
Vol.6 No.4,
November
13,
2017
ABSTRACT: Missense mutations in the leucine rich repeat kinase 2 (LRRK2) gene are the
leading genetic cause of autosomal dominant familial Parkinson’s disease. We
previously reported that two mutations within the ROC domain, namely
R1441C and A1442P, exhibit increased protein degradation leading to lowered
steady state LRRK2 protein levels in HEK293 cells. More recently, the common
WD40 domain LRRK2 haplotype, Met2397, which is a risk factor for
Crohn’s disease, has been shown to lower steady state protein levels in
HEK293 cells. In view of recent evidence implicating LRRK2 and inflamemation
in PD, we investigated the effects of Met2397 on LRRK2 expression,
and compared them to the Thr2397 variant and other LRRK2 mutants. In this
study, we transfected HEK293 cells with plasmid constructs encoding the different
LRRK2 variants, and analyzed the resulting protein levels by Western
blot and flow cytometry. Here we found that both the Met2397 and Thr2397
haplotypes yield similar levels of LRRK2 protein expression and do not appear
to impact cell viability in HEK293 cells, compared to other LRRK mutants.
Thus, we have concluded that the Met2397 haplotype is unlikely to play a role
in LRRK2 mediated or idiopathic PD.