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@ARTICLE{Gilsbach:274060,
author = {Gilsbach, Bernd K and Ho, Franz Y and Riebenbauer, Benjamin
and Zhang, Xiaojuan and Guaitoli, Giambattista and Kortholt,
Arjan and Gloeckner, Christian Johannes},
title = {{I}ntramolecular feedback regulation of the {LRRK}2 {R}oc
{G} domain by a {LRRK}2 kinase-dependent mechanism.},
journal = {eLife},
volume = {12},
issn = {2050-084X},
address = {Cambridge},
publisher = {eLife Sciences Publications},
reportid = {DZNE-2025-00041},
pages = {RP91083},
year = {2024},
abstract = {The Parkinson's disease (PD)-linked protein Leucine-Rich
Repeat Kinase 2 (LRRK2) consists of seven domains, including
a kinase and a Roc G domain. Despite the availability of
several high-resolution structures, the dynamic regulation
of its unique intramolecular domain stack is nevertheless
still not well understood. By in-depth biochemical analysis,
assessing the Michaelis-Menten kinetics of the Roc G domain,
we have confirmed that LRRK2 has, similar to other Roco
protein family members, a KM value of LRRK2 that lies within
the range of the physiological GTP concentrations within the
cell. Furthermore, the R1441G PD variant located within a
mutational hotspot in the Roc domain showed an increased
catalytic efficiency. In contrast, the most common PD
variant G2019S, located in the kinase domain, showed an
increased KM and reduced catalytic efficiency, suggesting a
negative feedback mechanism from the kinase domain to the G
domain. Autophosphorylation of the G1+2 residue (T1343) in
the Roc P-loop motif is critical for this phosphoregulation
of both the KM and the kcat values of the Roc-catalyzed GTP
hydrolysis, most likely by changing the monomer-dimer
equilibrium. The LRRK2 T1343A variant has a similar
increased kinase activity in cells compared to G2019S and
the double mutant T1343A/G2019S has no further increased
activity, suggesting that T1343 is crucial for the negative
feedback in the LRRK2 signaling cascade. Together, our data
reveal a novel intramolecular feedback regulation of the
LRRK2 Roc G domain by a LRRK2 kinase-dependent mechanism.
Interestingly, PD mutants differently change the kinetics of
the GTPase cycle, which might in part explain the difference
in penetrance of these mutations in PD patients.},
keywords = {Leucine-Rich Repeat Serine-Threonine Protein Kinase-2:
metabolism / Leucine-Rich Repeat Serine-Threonine Protein
Kinase-2: genetics / Leucine-Rich Repeat Serine-Threonine
Protein Kinase-2: chemistry / Humans / Phosphorylation /
Guanosine Triphosphate: metabolism / Kinetics / Protein
Domains / Parkinson Disease: genetics / Parkinson Disease:
metabolism / Feedback, Physiological / GTPase (Other) /
LRRK2 (Other) / Michaelis–Menten kinetics (Other) / PD
(Other) / biochemistry (Other) / chemical biology (Other) /
negative feedback loop (Other) / none (Other) / parkinson's
disease (Other) / Leucine-Rich Repeat Serine-Threonine
Protein Kinase-2 (NLM Chemicals) / LRRK2 protein, human (NLM
Chemicals) / Guanosine Triphosphate (NLM Chemicals)},
cin = {AG Gloeckner / AG Gasser},
ddc = {600},
cid = {I:(DE-2719)1210007 / I:(DE-2719)1210000},
pnm = {352 - Disease Mechanisms (POF4-352) / 353 - Clinical and
Health Care Research (POF4-353)},
pid = {G:(DE-HGF)POF4-352 / G:(DE-HGF)POF4-353},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39699947},
pmc = {pmc:PMC11658767},
doi = {10.7554/eLife.91083},
url = {https://pub.dzne.de/record/274060},
}