Structure of the Roc–COR domain tandem of C. tepidum, a prokaryotic homologue of the human LRRK2 Parkinson kinase

Gotthardt, Katja; Weyand, Michael; Kortholt, Arjan; Haastert, Peter J.M. van; Wittinghofer, Alfred

doi:10.1038/emboj.2008.150

Cited by 136 publications

(318 citation statements)

References 47 publications

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“…The existence of Roc ext as both dimers and monomers in solution indicate that it does not form constitutive dimer as previously suggested (10,11). This gives rise to the possibility of a dynamic nucleotide-dependent dimerization as a regulation mechanism of Roc activity suggested by Gotthardt et al (11).…”

Section: Significancementioning

confidence: 57%

“…The literature currently suggests that dimerization of Roc is required for its GTPase activity (10,11), but this requirement is uncertain because one of the two structures that provided this insight showed an unusual domain-swapped conformation not seen in other GTPases, whereas the second structure required intermolecular exchange of an arginine residue in trans to constitute an active site, but the required arginine does not exist in human LRRK2 (11). Having created a stable monomeric Roc construct enabled us to test these requirements.…”

Section: Significancementioning

confidence: 99%

“…This gives rise to the possibility of a dynamic nucleotide-dependent dimerization as a regulation mechanism of Roc activity suggested by Gotthardt et al (11). To investigate this possibility, we determined whether Roc 48 changes its conformation upon binding to guanine nucleotides by incubating Roc 48 with GDP or GppNHp for 2 h using standard nucleotide-exchange procedures (12).…”

Section: Significancementioning

confidence: 99%

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Parkinson disease-associated mutation R1441H in LRRK2 prolongs the “active state” of its GTPase domain

Liao

Burlak

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

106

136

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Mutation in leucine-rich-repeat kinase 2 (LRRK2) is a common cause of Parkinson disease (PD). A disease-causing point mutation R1441H/G/C in the GTPase domain of LRRK2 leads to overactivation of its kinase domain. However, the mechanism by which this mutation alters the normal function of its GTPase domain [Ras of complex proteins (Roc)] remains unclear. Here, we report the effects of R1441H mutation (Roc R1441H ) on the structure and activity of Roc. We show that Roc forms a stable monomeric conformation in solution that is catalytically active, thus demonstrating that LRRK2 is a bona fide self-contained GTPase. We further show that the R1441H mutation causes a twofold reduction in GTPase activity without affecting the structure, thermal stability, and GDP-binding affinity of Roc. However, the mutation causes a twofold increase in GTP-binding affinity of Roc, thus suggesting that the PD-causing mutation R1441H traps Roc in a more persistently activated state by increasing its affinity for GTP and, at the same time, compromising its GTP hydrolysis.M utation in leucine-rich-repeat kinase 2 (LRRK2) is a common cause of Parkinson disease (PD) (1-5). LRRK2 is a large (2,527-aa) multidomain protein consisting of seven putative domains (2), including a Ras-like GTPase domain called Ras of complex proteins (Roc), followed by a domain called C-terminal of Roc (COR), which is then followed by a kinase domain (Kin). It remains unclear how perturbations of these activities result in disease; however, the most common mutation in LRRK2-associated PD, G2019S in the kinase domain, shows higher kinase activity than wild type; therefore, its overactivation might be associated with disease pathogenesis (6).The tandem Roc-COR-Kin arrangement suggests that their activities might be coupled such that the GTPase activity of Roc might modulate the kinase activity. Indeed, several studies have shown that GTP binding to the Roc domain regulates the activity of the Kin domain (7,8). Moreover, a PD-associated mutation in the Roc domain (R1441C) has been shown to have higher kinase activity (9), thus suggesting that mutations in the Roc domain, also up-regulate kinase activity.Understanding the function of Roc and its mechanism of action is important for understanding the mechanism of PD pathogenesis and therapeutic development. However, because of the lack sufficient quantity of protein samples amendable for detailed investigations, the biochemical properties and enzymatic activities of the Roc domain of LRRK2 are poorly understood.Here, we describe a stably folded construct of human Roc domain that enabled us to investigate quantitatively its biochemical and enzymatic properties. The results revealed that a PD-causing mutation R1441H in the Roc domain renders it less active at hydrolyzing GTP, as well as having higher affinity for GTP, than its wild-type counterpart, thereby increasing the residence time of its GTP-bound "active state," which is associated with PD pathogenesis (8). Results and DiscussionConstruction of a Stable Human LRR...

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Section: Significancementioning

confidence: 57%

Section: Significancementioning

confidence: 99%

Section: Significancementioning

confidence: 99%

See 1 more Smart Citation

Parkinson disease-associated mutation R1441H in LRRK2 prolongs the “active state” of its GTPase domain

Liao

Burlak

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

106

136

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“…Dimerization is a common mechanism to regulate kinase activity (23). LRRK2, and most likely all Roco proteins, are dimer in solutions using the COR domain as dimerization interface (12,13,27). Autophosphorylation is essential for the function of LRRK2; however, data for autophosphorylation both in cis and in trans have been reported (15,27).…”

Section: Resultsmentioning

confidence: 97%

“…RocCOR proteins thus seem to belong to the G proteins activated by the nucleotide-dependent dimerization (GAD) class of molecular switches. PD-analogous mutations in Roc and COR alter the Roc-COR interface and result in decreased GTPase activity (12,13). The structure of the Roc domain of human LRRK2 showed a domainswapped dimeric G domain whose significance for the native protein is unclear (11,13).…”

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confidence: 99%

Roco kinase structures give insights into the mechanism of Parkinson disease-related leucine-rich-repeat kinase 2 mutations

Gilsbach

Vetter

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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127

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Mutations in human leucine-rich-repeat kinase 2 (LRRK2) have been found to be the most frequent cause of late-onset Parkinson disease. Here we show that Dictyostelium discoideum Roco4 is a suitable model to study the structural and biochemical characteristics of the LRRK2 kinase and can be used for optimization of current and identification of new LRRK2 inhibitors. We have solved the structure of Roco4 kinase wild-type, Parkinson disease-related mutants G1179S and L1180T (G2019S and I2020T in LRRK2) and the structure of Roco4 kinase in complex with the LRRK2 inhibitor H1152. Taken together, our data give important insight in the LRRK2 activation mechanism and, most importantly, explain the G2019S-related increase in LRRK2 kinase activity.

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LRRK2 and Parkinson Disease

Dächsel¹,

Farrer²

2010

Arch Neurol

138

114

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To review the molecular genetics and functional biology of leucine-rich repeat kinase 2 (LRRK2) in parkinsonism and to summarize the opportunities and challenges to develop interventions for Parkinson disease (PD) based on this genetic insight. Data Sources: Publications cited are focused on LRRK2 biology between 2004 and March 2009. Study Selection: Literature selected was based on original contributions, seminal observations, and thoughtful reviews. Data Extraction: Unless stated otherwise, data was primarily abstracted from peer-reviewed literature appearing on PubMed. Data Synthesis: Genetic mutations that predispose PD are diagnostically useful in early or atypical presentations. The molecular pathways identified suggest therapeutic interventions for Lrrk2 and idiopathic PD and the rationale and opportunity to develop physiologically relevant biomarkers and experimental models with which to test them. Conclusions: Both affected and asymptomatic LRRK2 carriers now provide the opportunity to define the natural history of PD. This includes the frequency, penetrance, and rate of motor symptoms, nonmotor comorbidities, and their associated biomarkers.

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Structure of the Roc–COR domain tandem of C. tepidum, a prokaryotic homologue of the human LRRK2 Parkinson kinase

Cited by 136 publications

References 47 publications

Parkinson disease-associated mutation R1441H in LRRK2 prolongs the “active state” of its GTPase domain

Parkinson disease-associated mutation R1441H in LRRK2 prolongs the “active state” of its GTPase domain

Roco kinase structures give insights into the mechanism of Parkinson disease-related leucine-rich-repeat kinase 2 mutations

LRRK2 and Parkinson Disease

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