The Parkinson disease causing LRRK2 mutation I2020T is associated with increased kinase activity.
Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) have been recently identified in families with autosomal dominant late-onset Parkinson disease (PD). The LRRK2 protein consists of multiple domains and belongs to the Roco family, a novel group of the Ras/GTPase superfamily. Besides the GTPase (Roc) domain, it contains a predicted kinase domain, with homology to MAP kinase kinase kinases. Using cell fractionation and immunofluorescence microscopy, we show that LRRK2 is localized in the cytoplasm and is associated with cellular membrane structures. The purified LRRK2 protein demonstrates autokinase activity. The disease-associated I2020T mutant shows a significant increase in autophosphorylation of approximately 40% in comparison to wild-type protein in vitro. This suggests that the pathology of PD caused by the I2020T mutation is associated with an increase rather than a loss in LRRK2 kinase activity.
Gloeckner
Christian Johannes
CJ
GSF-National Research Center for Environment and Health, Institute of Human Genetics, Munich-Neuherberg, Germany.
Kinkl
Norbert
N
Schumacher
Annette
A
Braun
Ralf J
RJ
O'Neill
Eric
E
Meitinger
Thomas
T
Kolch
Walter
W
Prokisch
Holger
H
Ueffing
Marius
M
D015153:Blotting, Western; D002458:Cell Fractionation; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D020125:Mutation, Missense / Q000235:genetics*; D010300:Parkinson Disease / Q000201:enzymology* / Q000235:genetics*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*
C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases