9CHO

Autoinhibited full-length LRRK2(I2020T) on microtubules with MLi-2

Summary for 9CHO

• Entry DOI: 10.2210/pdb9cho/pdb
• EMDB information: 45591 45593 45594 45595 45596
• Descriptor: Leucine-rich repeat serine/threonine-protein kinase 2, GUANOSINE-5'-DIPHOSPHATE, (2~{R},6~{S})-2,6-dimethyl-4-[6-[5-(1-methylcyclopropyl)oxy-1~{H}-indazol-3-yl]pyrimidin-4-yl]morpholine (3 entities in total)
• Functional Keywords: kinase, gtpase, protein binding
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 225727.53

Authors

Chen, S.,Villa, E.,Leschziner, A.E. (deposition date: 2024-07-01, release date: 2024-12-25, Last modification date: 2025-01-01)

Primary citation

Chen, S.,Basiashvili, T.,Hutchings, J.,Murillo, M.S.,Suarez, A.V.,Louro, J.A.,Leschziner, A.E.,Villa, E.
Cryo-electron tomography reveals the microtubule-bound form of inactive LRRK2.
Biorxiv, 2024

PubMed Abstract: Parkinson's Disease (PD) is the second most common neurodegenerative disorder. Mutations in leucine-rich repeat kinase 2 (LRRK2), a multi-domain protein containing both a kinase and a GTPase, are a leading cause of the familial form of PD. Pathogenic LRRK2 mutations increase LRRK2 kinase activity. While the bulk of LRRK2 is found in the cytosol, the protein associates with membranes where its Rab GTPase substrates are found, and under certain conditions, with microtubules. Integrative structural studies using single-particle cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) have revealed the architecture of microtubule-associated LRRK2 filaments, and that formation of these filaments requires LRRK2's kinase to be in the active-like conformation. However, whether LRRK2 can interact with and form filaments on microtubules in its autoinhibited state, where the kinase domain is in the inactive conformation and the N-terminal LRR domain covers the kinase active site, was not known. Using cryo-ET, we show that full-length LRRK2 can oligomerize on microtubules in its autoinhibited state. Both WT-LRRK2 and PD-linked LRRK2 mutants formed filaments on microtubules. While these filaments are stabilized by the same interfaces seen in the active-LRRK2 filaments, we observed a new interface involving the N-terminal repeats that were disordered in the active-LRRK2 filaments. The helical parameters of the autoinhibited-LRRK2 filaments are different from those reported for the active-LRRK2 filaments. Finally, the autoinhibited-LRRK2 filaments are shorter and less regular, suggesting they are less stable.

PubMed: 38948781
DOI: 10.1101/2024.06.18.599606

Experimental method

ELECTRON MICROSCOPY (7.8 Å)