Parkinson's disease-associated mutations in the GTPase domain of LRRK2 impair its nucleotide-dependent conformational dynamics

Wu, Chunxiang; Liao, Jingling; Park, Yangshin; Reed, Xylena; Engel, Victoria A.; Hoang, Neo C.; Takagi, Yuichiro; Johnson, Steven M.; Wang, Mu; Federici, Mark G.; Nichols, R. Jeremy; Sanishvili, Ruslan; Cookson, Mark; Hoang, Quyen Q.

doi:10.1074/jbc.ra119.007631

Cited by 26 publications

(32 citation statements)

References 22 publications

(25 reference statements)

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“…The architecture of the ROC domain of LRRK2 While a structure of the ROC domain of LRRK2 has been determined a decade ago (PDB ID: 2ZEJ) (17), that structure has been inconsistent with subsequent biochemical and biophysical observations. For example, the 2ZEJ structure showed an obligate dimer with an active-site composed of one half from each protomer; however, biochemical studies have demonstrated that ROC dynamically interconverts between its dimeric and monomeric conformations and that the monomers are catalytically active (14,15). Thus, the examination of the 2ZEJ structure did not provide us with the insights that we sought to understand the conformational dynamics ROC and the mechanisms of the PD-associated mutations.…”

Section: Resultsmentioning

confidence: 99%

“…For example, the PD-associated residues in HsROC are not conserved in CtROC, including residue R1441 in HsROC; therefore, the potential biochemical and structural effects of the PD-associated mutations cannot be inferred directly from the CtROC structures. Due to these reasons, we set out to determine the structure of the human ROC ext construct to examine the structural bases for the biochemical and conformational dynamics we observed (14,15,20).…”

Section: Resultsmentioning

confidence: 99%

“…Subsequently, we showed that GTP-binding drives dimeric ROC ext disassembly into monomers and GDP-binding oppositely shifts the equilibrium back to dimers (15). Moreover, we showed that all the PD-associated mutations at Arg-1441, as well as N1437H, impair the dimermonomer dynamics of ROC ext (15,16).…”

Section: Significance Statementmentioning

confidence: 87%

“…Indeed, we recently engineered a stable construct of ROC (ROC ext ) and showed that the R1441H mutation perturbs the hydrolytic conversion of GTP to GDP, thereby prolonging the GTP-bound "on" state of ROC (14). Subsequently, we showed that GTP-binding drives dimeric ROC ext disassembly into monomers and GDP-binding oppositely shifts the equilibrium back to dimers (15). Moreover, we showed that all the PD-associated mutations at Arg-1441, as well as N1437H, impair the dimermonomer dynamics of ROC ext (15,16).…”

Section: Significance Statementmentioning

confidence: 99%

“…Second, the disease-associated residues R1441, N1437, and R1398 in HsROC structurally aligned with residues Y558, H554, and Q519 of CtROC, respectively. The difference in amino acids at these positions is important in that the substitution of residue Arg1441 in HsROC for Tyr found in the equivalent position in CtROC, and residue N1437 in HsROC for His of CtROC both resulted in impairment of the conformational dynamics of ROC ext (15,16) (Fig. 3c).…”

Section: Comparison Of Human Roc With C Tepidum Rocomentioning

confidence: 99%

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A revised 1.6 Å structure of the GTPase domain of the Parkinson’s disease-associated protein LRRK2 provides insights into mechanisms

Liao

Park

et al. 2019

Preprint

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Leucine-rich repeat kinase 2 (LRRK2) is a large 286 kDa multi-domain protein whose mutation is a common cause of Parkinson’s disease (PD). One of the common sites of familial PD-associated mutations occurs at residue Arg-1441 in the GTPase domain of LRRK2. Previously, we reported that the PD-associated mutation R1441H impairs the catalytic activity of the GTPase domain thereby traps it in a persistently "on" state. More recently, we reported that the GTPase domain of LRRK2 exists in a dynamic dimer-monomer equilibrium where GTP binding shifts it to the monomeric conformation while GDP binding shifts it back to the dimeric state. We also reported that all of the PD-associated mutations at Arg-1441, including R1441H, R1441C, and R1441G, impair the nucleotide-dependent dimer-monomer conformational dynamics of the GTPase domain. However, the mechanism of this nucleotide-dependent conformational dynamics and how it is impaired by the mutations at residue Arg-1441 remained unclear. Here, we report a 1.6 Å crystal structure of the GTPase domain of LRRK2. Our structure has revealed a dynamic switch region that can be differentially regulated by GTP and GDP binding. This nucleotide-dependent regulation is impaired when residue Arg-1441 is substituted with the PD-associated mutations due to the loss of its exquisite interactions consisting of two hydrogen bonds and a π-stacking interaction at the dimer interface.Significance StatementMutations in LRRK2 are associated with familial Parkinson’s disease, so understanding its mechanism of actions and how they are changed by the disease-associated mutations is important for developing therapeutic strategies. This paper describes an atomic structure of the G-domain of LRRK2 revealing that the conformational dynamics of the switch regions are potentially important for its normal function. It further shows that a disease-associated mutation could lock the G domain in a persistently active-like conformation, thus perturbing its normal function.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%