Intramolecular feedback regulation of the LRRK2 Roc G domain by a LRRK2 kinase dependent mechanism

• Published in: bioRxiv
• Main Authors: Gilsbach, Bernd K, Ho, Franz Y, Riebenbauer, Benjamin, Zhang, Xiaojuan, Guaitoli, Giambattista, Kortholt, Arjan, Gloeckner, Christian Johannes
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 27.02.2024
• Subjects: Biochemical analysis; Feedback; Kinases; LRRK2 protein; Movement disorders; Mutation hot spots; Neurodegenerative diseases; Parkinson's disease; Phosphorylation; Statistical analysis
• DOI: 10.1101/2023.07.31.549909

The Parkinson's Disease (PD)-linked protein Leucine Rich Repeat Kinase 2 (LRRK2) consists of seven domains, including a kinase and a Roc G domain. Despite the availability of several high-resolution structures, the dynamic regulation of its unique intramolecular domain stack is nevertheless still not well understood. By in-depth biochemical analysis, assessing the Michaelis-Menten kinetics of the Roc G domain, we have confirmed that LRRK2 has similar to other Roco protein family members a KM value of LRRK2 that lays within the range of the physiological GTP concentrations within the cell. Furthermore, the R1441G PD variant located within a mutational hotspot in the Roc domain showed an increased catalytic efficiency. The most common PD variant G2019S, located in the kinase domain showed an increased KM and reduced catalytic efficiency, suggesting a negative feedback mechanism from the kinase domain to the G-domain. Auto-phosphorylation of the G1+2 residue (T1343) in the Roc P-loop motif is critical for this phosphoregulation of both the KM as well as the kcat values of the Roc-catalyzed GTP hydrolysis, most likely by changing the monomer-dimer equilibrium. Together our data reveal a novel intramolecular feedback regulation of the LRRK2 Roc G domain by a LRRK2 kinase dependent mechanism. Interestingly, PD mutants differently change the kinetics of the GTPase cycle, which might in part explain the difference in penetrance of these mutations in PD patients.Competing Interest StatementThe authors have declared no competing interest.Footnotes* Revision to address the reviews: - Changes throughout the text. - Revised Figure 5, Figure 6 and Supplemental Figure 4. - Supplemental data on statistical analyses were provided.