Structural basis of human LRRK2 membrane recruitment and activation

Abstract

SummaryMutations in LRRK2 are the most common genetic cause of late-onset Parkinson’s disease (PD). LRRK2 encodes the leucine-rich repeat kinase 2 (LRRK2), whose kinase activity is regulated by Rab29, a membrane-anchored GTPase. However, molecular mechanisms underlying Rab29-dependent recruitment and activation of LRRK2 remain unclear. Here we report cryo-EM structures of LRRK2–Rab29 complexes in three oligomeric states, illustrating snapshots of key steps during LRRK2 membrane recruitment and activation. Rab29 binds to the ARM domain of LRRK2, and disruption at the interface abrogates LRRK2 kinase activity. Activation of LRRK2 is underpinned by the formation of an unexpected Rab29-induced super-assembly containing two central kinase-active and two peripheral kinase-inactive LRRK2 protomers. Central protomers undergo pronounced oligomerization-associated rearrangements and adopt an active conformation. Our work reveals the structural mechanism for LRRK2’s spatial regulation controlled by Rab GTPases, provides mechanistic insights into pathogenic mutations and identifies new opportunities to design LRRK2 inhibitors for PD treatment.

PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORBGTPase domain

“…The unstructured nature of the loop makes it difficult to predict how phosphorylation the COR B loop impacts folding and interaction with the kinase αC-helix. Recently, a 3.5 Å structure of an active tetrameric form of LRRK2 has been reported [18]. This consists of two peripheral LRRK2 molecules in the inactive conformation and two central LRRK2 protomers in an active conformation [18].…”

“…Recently, a 3.5 Å structure of an active tetrameric form of LRRK2 has been reported [18]. This consists of two peripheral LRRK2 molecules in the inactive conformation and two central LRRK2 protomers in an active conformation [18]. The residues in LRRK2 that lie at the equivalent region to Ser1064, Ser1074 and Thr1075 in the active LRRK2 subunits encompass residues 1721-1725 in LRRK2 (Fig 9, Fig 6C).…”

“…Our previous work suggests that LRRK1, but not LRRK2, is stimulated by phorbol esters that switch on conventional and novel PKC isoforms [8]. In contrast, LRRK2 is activated by recruitment to membrane vesicles, through binding to Rab GTPases such as Rab29 that interacts with the ARM domain that is not present in LRRK1 [14][15][16][17][18]. Furthermore, a Parkinson's causing mutation in VPS35[D620N] stimulates LRRK2 activity without impacting LRRK1 [8,19].…”

Impact of 100 LRRK2 variants linked to Parkinson’s Disease on kinase activity and microtubule binding

“…LRRK2 is activated following its recruitment to cellular membranes via interactions with Rab29 and other Rab GTPases, likely via its N-terminal ARM domain [11–15]. LRRK2 phosphorylates a subgroup of Rab GTPases at membranes, including Rab8A and Rab10, at a conserved Ser/Thr residue located within the effector binding Switch-II domain [16–18].…”

Endogenous Rab38 regulates LRRK2’s membrane recruitment and substrate Rab phosphorylation in melanocytes

“…12, 13 Further, Rab29 protein and LRRK2 physically interact, with Rab29 directly binding LRRK2’s ARM domain, particularly LRRK2 360-450 . 14–16 Very recent work also identified the extreme N-terminus of LRRK2 as a high-affinity binding site for LRRK2-phosphorylated Rabs, suggesting a feed-forward pathway causing LRRK2 and phospho-Rab accumulation at membranes. 15 In the cell, overexpressed Rab29 drives overexpressed LRRK2 kinase activation by increasing membrane localization of LRRK2.…”