Roco Proteins and the Parkinson’s Disease-Associated LRRK2

Liao, Jingling; Hoang, Quyen Q.

doi:10.3390/ijms19124074

Cited by 8 publications

(7 citation statements)

References 95 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover, mutations in CtRoco at Tyr-558 mimicking the LRRK2 R1441G/R1441C/ R1441H did not affect its dimerization, whereas these mutations in LRRK2 ROC ext abolished dimer formation as shown above. These similarities and differences between CtRoco and LRRK2 suggest that the subtle variability in the mechanisms of action observed in the typical Ras and Rab families of GTPases might also occur in the Roco family of GTPases (19).…”

Section: Mutations In Roc Of Lrrk2 Impair Conformational Dynamicsmentioning

confidence: 99%

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

Liao

Park

et al. 2019

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Section: Mutations In Roc Of Lrrk2 Impair Conformational Dynamicsmentioning

confidence: 99%

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

Liao

Park

et al. 2019

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

“…LRRK2 belongs to the protein family of ROCO proteins, which are defined by containing a GTPase domain called Ras of Complex (Roc) followed by a COR domain in combination with other domains. A special feature of LRRK2, as well as some but not all of the ROC:COR proteins, is that it contains both, a kinase domain and a GTPase domain so that two catalytically active core domains are embedded within the same polypeptide chain (9)(10)(11).…”

Section: Introductionmentioning

confidence: 99%

Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2

Schmidt

Weng²,

Aoto³

et al. 2020

Preprint

View full text Add to dashboard Cite

In a multi-tiered approach, we explored how Parkinson's Disease-related mutations hijack the finely tuned activation process of Leucine-Rich Repeat Kinase 2 (LRRK2) using a construct containing the ROC, Cor, Kinase and WD40 domains (LRRK2RCKW). We hypothesized that the N-terminal domains shield the catalytic domains in an inactive state. PD mutations, type-I LRRK2 inhibitors, or physiological Rab GTPases can unleash the catalytic domains while the active kinase conformation, but not kinase activity, is essential for docking onto microtubules. Mapping solvent accessible regions of LRRK2RCKW employing hydrogen-deuterium exchange mass spectrometry (HDX-MS) revealed how inhibitor binding is sensed by the entire protein. Molecular Dynamics simulations of the kinase domain elucidated differences in conformational dynamics between wt and mutants of the DYGψ motif. While all domains contribute to regulating kinase activity and spatial distribution, the kinase domain, driven by the DYGψ motif, coordinates domain crosstalk and serves as an intrinsic hub for LRRK2 regulation.

show abstract

“…LRRK2 belongs to the protein family of ROCO proteins, which are de ned by containing a GTPase domain called Ras of Complex (ROC) followed by a COR domain in combination with other domains. A special feature of LRRK2, as well as some but not all of the ROC:COR proteins, is that it contains both, a kinase domain and a GTPase domain so that two catalytically active core domains are embedded within the same polypeptide chain (9)(10)(11).…”

Section: Introductionmentioning

confidence: 99%

Conformation and Dynamics of the Kinase Domain Drive Subcellular Location and Activation of LRRK2.

Schmidt

Weng

Aoto

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Leucine-Rich Repeat Kinase 2 (LRRK2) is a complex multi-domain protein where LRRK2-mutations are associated with Parkinson´s Disease (PD). To explore how pathogenic PD-mutations hijack the finely tuned activation process of LRRK2, we here used a multi-tiered approach. Methods: First, the spatial and temporal distribution of full-length LRRK2 was investigated by a real-time cell-based assay in the presence and absence of LRRK2-kinase inhibitors. In a 2nd layer we explored the consequences of PD mutations as well as removal of the N-terminal domains employing a construct containing the ROC, Cor, Kinase and WD40 domains (LRRK2RCKW). We focused on the biochemical characterization of LRRK2RCKW variants based on kinase assays using Rab8a or LRRKtide as substrates. Next, we used hydrogen-deuterium exchange mass spectrometry (HDX-MS) to map the solvent accessible regions of LRRK2RCKW in the presence and absence of the LRRK2 inhibitor MLi-2. Finally, Molecular Dynamics simulations on the kinase domain were applied to elucidate differences in breathing dynamics between wild type and mutants of the DYGψ motif. Results: Our cellular approaches revealed that the kinase inhibitors MLi-2 and rebastinib both freeze the kinase domain in a stable conformation, however, only MLi-2 resembles an active conformation and induces filament formation. LRRK2RCKW showed, regardless of the mutation it was combined with, filament formation, indicating a shielding function of the N-terminal domains. This shielding function is impaired for pathogenic mutations in full length LRRK2. LRRK2RCKW retained kinase activity similar to full-length LRRK2. HDX-MS provided a comprehensive allosteric portrait of the kinase domain and revealed how MLi-2 binding is sensed by the entire protein. Molecular Dynamics simulations suggest that, while all domains contribute to regulating kinase activity and spatial distribution, it is the highly dynamic kinase domain, driven by the DYGψ motif, that coordinates the overall domain crosstalk and serves as a regulatory hub for the intrinsic regulation of LRRK2.Conclusion: These studies confirm our hypothesis that the N-terminal scaffolding domains shield the catalytic domains in an inactive state. PD mutations, MLi-2, or Rab GTPases can all unleash the catalytic domains while the active kinase conformation, but not kinase activity, is essential for docking onto microtubules.

show abstract

Roco Proteins and the Parkinson’s Disease-Associated LRRK2

Cited by 8 publications

References 95 publications

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

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

Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2

Conformation and Dynamics of the Kinase Domain Drive Subcellular Location and Activation of LRRK2.

Contact Info

Product

Resources

About