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Cited by 0Parkinson's Disease Mechanisms and TreatmentsProtein Kinase Regulation and GTPase SignalingReceptor Mechanisms and Signaling

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Structural insights into the GTP-driven monomerization and activation of a bacterial LRRK2 homolog using allosteric nanobodies

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Galicia, Christian, Guaitoli, Giambattista, Fislage, Marcus, Gloeckner, Christian Johannes, Versées, Wim·2024·eLife
DOI 10.7554/elife.94503.3

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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: Nb Roco1 and Nb Roco2 . 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 Nb Roco1 and Nb Roco2 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.

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Metadata

Authors
Galicia, C., Guaitoli, G., Fislage, M., Gloeckner, C. J., & Versées, W.
Affiliations
  • Vrije Universiteit Brussel (BE)
  • German Center for Neurodegenerative Diseases (DE)
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2024-04-26
Volume
13
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.94503.3
OpenAlex
W4395667074
Cited by
0
References
3
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
eLife (publishedVersion, cc-by)
Funders
  • Fonds Wetenschappelijk Onderzoek (G005219N, G003322N)
  • Vrije Universiteit Brussel (SRP50, SRP95)
Topics
  • Health Sciences › Medicine › Clinical Neurology › Parkinson's Disease Mechanisms and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
LRRK2, GTPase activation, allosteric activation, nanobodies, GTP-bound state, protein monomerization, Parkinson's disease, GTPase domain, allosteric modulation

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