TY  - JOUR
AU  - Galicia, Christian
AU  - Guaitoli, Giambattista
AU  - Fislage, Marcus
AU  - Gloeckner, Christian Johannes
AU  - Versées, Wim
TI  - Structural insights into the GTP-driven monomerization and activation of a bacterial LRRK2 homolog using allosteric nanobodies.
JO  - eLife
VL  - 13
SN  - 2050-084X
CY  - Cambridge
PB  - eLife Sciences Publications
M1  - DZNE-2024-00514
SP  - RP94503
PY  - 2024
AB  - 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.
KW  - Cryoelectron Microscopy
KW  - Single-Domain Antibodies: metabolism
KW  - Single-Domain Antibodies: chemistry
KW  - Guanosine Triphosphate: metabolism
KW  - Guanosine Triphosphate: chemistry
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2: metabolism
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2: chemistry
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2: genetics
KW  - Bacterial Proteins: metabolism
KW  - Bacterial Proteins: chemistry
KW  - Bacterial Proteins: genetics
KW  - Protein Conformation
KW  - Allosteric Regulation
KW  - Models, Molecular
KW  - Protein Multimerization
KW  - Humans
KW  - LRRK2 (Other)
KW  - Parkinson's disease (Other)
KW  - allosteric mechanism (Other)
KW  - cryo-EM (Other)
KW  - molecular biophysics (Other)
KW  - nanobodies (Other)
KW  - none (Other)
KW  - structural biology (Other)
KW  - Single-Domain Antibodies (NLM Chemicals)
KW  - Guanosine Triphosphate (NLM Chemicals)
KW  - Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (NLM Chemicals)
KW  - Bacterial Proteins (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:38666771
C2  - pmc:PMC11052575
DO  - DOI:10.7554/eLife.94503
UR  - https://pub.dzne.de/record/269345
ER  -