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Structural biology of the LRRK2 GTPase and kinase domains: implications for regulation.

Gilsbach BK, Kortholt A - Front Mol Neurosci (2014)

Bottom Line: Several of the pathogenic mutations in LRRK2 result in decreased GTPase activity and enhanced kinase activity, suggesting a possible PD-related gain of abnormal function.Studies with Roco proteins from the model organism Dictyostelium discoideum revealed that PD mutants have different effects and most importantly they explained the G2019S-related increased LRRK2 kinase activity.In this review we highlight the recent progress in structural and biochemical characterization of Roco proteins and discuss its implication for the understanding of the complex regulatory mechanism of LRRK2.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biochemistry, University of Groningen Groningen, Netherlands.

ABSTRACT
Human leucine rich repeat kinase 2 (LRRK2) belongs to the Roco family of proteins, which are characterized by the presence of a Ras-like G-domain (Roc), a C-terminal of Roc domain (COR), and a kinase domain. Mutations in LRRK2 have been found to be thus far the most frequent cause of late-onset Parkinson's disease (PD). Several of the pathogenic mutations in LRRK2 result in decreased GTPase activity and enhanced kinase activity, suggesting a possible PD-related gain of abnormal function. Important progress in the structural understanding of LRRK2 has come from our work with related Roco proteins from lower organisms. Atomic structures of Roco proteins from prokaryotes revealed that Roco proteins belong to the GAD class of molecular switches (G proteins activated by nucleotide dependent dimerization). As in LRRK2, PD-analogous mutations in Roco proteins from bacteria decrease the GTPase reaction. Studies with Roco proteins from the model organism Dictyostelium discoideum revealed that PD mutants have different effects and most importantly they explained the G2019S-related increased LRRK2 kinase activity. Furthermore, the structure of Dictyostelium Roco4 kinase in complex with the LRRK2 inhibitor H1152 showed that Roco4 and other Roco family proteins can be important for the optimization of the current, and identification of new, LRRK2 kinase inhibitors. In this review we highlight the recent progress in structural and biochemical characterization of Roco proteins and discuss its implication for the understanding of the complex regulatory mechanism of LRRK2.

No MeSH data available.


Related in: MedlinePlus

Crystal structures of the human swapped Roc dimer and the C. tepidum RocCOR dimer. (A) Human Roc Dimer depicted as a cartoon with Roc-A in orange and Roc-B in red. Above the domain representation of LRRK2 is shown. (B) Domain representation of C. tepidum and below a cartoon representation of the C. tepidum RocCOR structure with RocCOR-A in green and COR-B in blue. (C) Overlay of the two structures Roc-A (orange) of the human protein clashes with the N-terminal part of the C. tepidum COR-A (green). [PDB: 2ZEJ (human Roc), 3DPU (C. tepidum RocCOR)].
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Figure 3: Crystal structures of the human swapped Roc dimer and the C. tepidum RocCOR dimer. (A) Human Roc Dimer depicted as a cartoon with Roc-A in orange and Roc-B in red. Above the domain representation of LRRK2 is shown. (B) Domain representation of C. tepidum and below a cartoon representation of the C. tepidum RocCOR structure with RocCOR-A in green and COR-B in blue. (C) Overlay of the two structures Roc-A (orange) of the human protein clashes with the N-terminal part of the C. tepidum COR-A (green). [PDB: 2ZEJ (human Roc), 3DPU (C. tepidum RocCOR)].

Mentions: In Roco family members, the G-domain always occurs in tandem with the COR domain. There are two crystal structures comprising the Roc G-domain available: one structure of the LRRK2 Roc domain and one of the Roc-COR tandem of the Roco protein from Chlorobium tepidum (Deng et al., 2008; Gotthardt et al., 2008). Surprisingly, the structure of the LRRK2 Roc domain revealed a swapped dimer: in which the N-terminal part of one G-domain interacts with the C-terminal of the other, thereby forming a constitutive dimer (Deng et al., 2008). In contrast, the Roc domain in the C. tepidum RocCOR dimer structure shows the typical small G protein fold with six β-strands and helices on both sides and an additional N-terminal helix, termed α0-helix (Figure 3). The COR domain consists of two parts: the highly conserved N-terminal part interacts with the Roc domain and the less conserved C-terminal part functions as dimerization device. It seems rather unlikely that the human RocCOR tandem has a different folding than that of the bacterial Roco protein. Importantly, an overlay of the human Roc and the bacterial RocCOR structure revealed major clashes of the highly conserved N-terminal part of the COR domain with the swapped Roc dimer [Figure 3, (Gotthardt et al., 2008)]. Furthermore, Deng et al. (2008) could not convincingly show dimer formation of the Roc domain in solution, while Liao et al. (2014) showed that human Roc forms primarily a monomer in solution with low GTPase activity. (Deng et al., 2008; Liao et al., 2014) Together, this strongly suggest that, like all previously observed swapped G-protein structures (Chavas et al., 2007), the LRRK2 Roc structure is a crystallographic artifact.


Structural biology of the LRRK2 GTPase and kinase domains: implications for regulation.

Gilsbach BK, Kortholt A - Front Mol Neurosci (2014)

Crystal structures of the human swapped Roc dimer and the C. tepidum RocCOR dimer. (A) Human Roc Dimer depicted as a cartoon with Roc-A in orange and Roc-B in red. Above the domain representation of LRRK2 is shown. (B) Domain representation of C. tepidum and below a cartoon representation of the C. tepidum RocCOR structure with RocCOR-A in green and COR-B in blue. (C) Overlay of the two structures Roc-A (orange) of the human protein clashes with the N-terminal part of the C. tepidum COR-A (green). [PDB: 2ZEJ (human Roc), 3DPU (C. tepidum RocCOR)].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4017136&req=5

Figure 3: Crystal structures of the human swapped Roc dimer and the C. tepidum RocCOR dimer. (A) Human Roc Dimer depicted as a cartoon with Roc-A in orange and Roc-B in red. Above the domain representation of LRRK2 is shown. (B) Domain representation of C. tepidum and below a cartoon representation of the C. tepidum RocCOR structure with RocCOR-A in green and COR-B in blue. (C) Overlay of the two structures Roc-A (orange) of the human protein clashes with the N-terminal part of the C. tepidum COR-A (green). [PDB: 2ZEJ (human Roc), 3DPU (C. tepidum RocCOR)].
Mentions: In Roco family members, the G-domain always occurs in tandem with the COR domain. There are two crystal structures comprising the Roc G-domain available: one structure of the LRRK2 Roc domain and one of the Roc-COR tandem of the Roco protein from Chlorobium tepidum (Deng et al., 2008; Gotthardt et al., 2008). Surprisingly, the structure of the LRRK2 Roc domain revealed a swapped dimer: in which the N-terminal part of one G-domain interacts with the C-terminal of the other, thereby forming a constitutive dimer (Deng et al., 2008). In contrast, the Roc domain in the C. tepidum RocCOR dimer structure shows the typical small G protein fold with six β-strands and helices on both sides and an additional N-terminal helix, termed α0-helix (Figure 3). The COR domain consists of two parts: the highly conserved N-terminal part interacts with the Roc domain and the less conserved C-terminal part functions as dimerization device. It seems rather unlikely that the human RocCOR tandem has a different folding than that of the bacterial Roco protein. Importantly, an overlay of the human Roc and the bacterial RocCOR structure revealed major clashes of the highly conserved N-terminal part of the COR domain with the swapped Roc dimer [Figure 3, (Gotthardt et al., 2008)]. Furthermore, Deng et al. (2008) could not convincingly show dimer formation of the Roc domain in solution, while Liao et al. (2014) showed that human Roc forms primarily a monomer in solution with low GTPase activity. (Deng et al., 2008; Liao et al., 2014) Together, this strongly suggest that, like all previously observed swapped G-protein structures (Chavas et al., 2007), the LRRK2 Roc structure is a crystallographic artifact.

Bottom Line: Several of the pathogenic mutations in LRRK2 result in decreased GTPase activity and enhanced kinase activity, suggesting a possible PD-related gain of abnormal function.Studies with Roco proteins from the model organism Dictyostelium discoideum revealed that PD mutants have different effects and most importantly they explained the G2019S-related increased LRRK2 kinase activity.In this review we highlight the recent progress in structural and biochemical characterization of Roco proteins and discuss its implication for the understanding of the complex regulatory mechanism of LRRK2.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biochemistry, University of Groningen Groningen, Netherlands.

ABSTRACT
Human leucine rich repeat kinase 2 (LRRK2) belongs to the Roco family of proteins, which are characterized by the presence of a Ras-like G-domain (Roc), a C-terminal of Roc domain (COR), and a kinase domain. Mutations in LRRK2 have been found to be thus far the most frequent cause of late-onset Parkinson's disease (PD). Several of the pathogenic mutations in LRRK2 result in decreased GTPase activity and enhanced kinase activity, suggesting a possible PD-related gain of abnormal function. Important progress in the structural understanding of LRRK2 has come from our work with related Roco proteins from lower organisms. Atomic structures of Roco proteins from prokaryotes revealed that Roco proteins belong to the GAD class of molecular switches (G proteins activated by nucleotide dependent dimerization). As in LRRK2, PD-analogous mutations in Roco proteins from bacteria decrease the GTPase reaction. Studies with Roco proteins from the model organism Dictyostelium discoideum revealed that PD mutants have different effects and most importantly they explained the G2019S-related increased LRRK2 kinase activity. Furthermore, the structure of Dictyostelium Roco4 kinase in complex with the LRRK2 inhibitor H1152 showed that Roco4 and other Roco family proteins can be important for the optimization of the current, and identification of new, LRRK2 kinase inhibitors. In this review we highlight the recent progress in structural and biochemical characterization of Roco proteins and discuss its implication for the understanding of the complex regulatory mechanism of LRRK2.

No MeSH data available.


Related in: MedlinePlus