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@ARTICLE{Galicia:269345,
author = {Galicia, Christian and Guaitoli, Giambattista and Fislage,
Marcus and Gloeckner, Christian Johannes and Versées, Wim},
title = {{S}tructural insights into the {GTP}-driven monomerization
and activation of a bacterial {LRRK}2 homolog using
allosteric nanobodies.},
journal = {eLife},
volume = {13},
issn = {2050-084X},
address = {Cambridge},
publisher = {eLife Sciences Publications},
reportid = {DZNE-2024-00514},
pages = {RP94503},
year = {2024},
abstract = {Roco proteins entered the limelight after mutations in
human LRRK2 were identified as a major cause of familial
Parkinson's disease. LRRK2 is a large and complex protein
combining a GTPase and protein kinase activity, and disease
mutations increase the kinase activity, while presumably
decreasing the GTPase activity. Although a
cross-communication between both catalytic activities has
been suggested, the underlying mechanisms and the regulatory
role of the GTPase domain remain unknown. Several structures
of LRRK2 have been reported, but structures of Roco proteins
in their activated GTP-bound state are lacking. Here, we use
single-particle cryo-electron microscopy to solve the
structure of a bacterial Roco protein (CtRoco) in its
GTP-bound state, aided by two conformation-specific
nanobodies: NbRoco1 and NbRoco2. This structure presents
CtRoco in an active monomeric state, featuring a very large
GTP-induced conformational change using the LRR-Roc linker
as a hinge. Furthermore, this structure shows how NbRoco1
and NbRoco2 collaborate to activate CtRoco in an allosteric
way. Altogether, our data provide important new insights
into the activation mechanism of Roco proteins, with
relevance to LRRK2 regulation, and suggest new routes for
the allosteric modulation of their GTPase activity.},
keywords = {Cryoelectron Microscopy / Single-Domain Antibodies:
metabolism / Single-Domain Antibodies: chemistry / Guanosine
Triphosphate: metabolism / Guanosine Triphosphate: chemistry
/ Leucine-Rich Repeat Serine-Threonine Protein Kinase-2:
metabolism / Leucine-Rich Repeat Serine-Threonine Protein
Kinase-2: chemistry / Leucine-Rich Repeat Serine-Threonine
Protein Kinase-2: genetics / Bacterial Proteins: metabolism
/ Bacterial Proteins: chemistry / Bacterial Proteins:
genetics / Protein Conformation / Allosteric Regulation /
Models, Molecular / Protein Multimerization / Humans / LRRK2
(Other) / Parkinson's disease (Other) / allosteric mechanism
(Other) / cryo-EM (Other) / molecular biophysics (Other) /
nanobodies (Other) / none (Other) / structural biology
(Other) / Single-Domain Antibodies (NLM Chemicals) /
Guanosine Triphosphate (NLM Chemicals) / Leucine-Rich Repeat
Serine-Threonine Protein Kinase-2 (NLM Chemicals) /
Bacterial Proteins (NLM Chemicals)},
cin = {AG Gloeckner},
ddc = {600},
cid = {I:(DE-2719)1210007},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:38666771},
pmc = {pmc:PMC11052575},
doi = {10.7554/eLife.94503},
url = {https://pub.dzne.de/record/269345},
}