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2024-04-03Author
Trilling, Chiara R.Weng, Jui-HungKaila Sharma, PallaviNolte, ViktoriaWu, JianMa, WenBoassa, DanielaTaylor, Susan T.Herberg, Friedrich WilhelmMetadata
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RedOx regulation of LRRK2 kinase activity by active site cysteines
Abstract
Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson’s disease (PD). Most of these pathogenic mutations are located in the kinase domain (KD) or GTPase domain of LRRK2. In this study we describe a mechanism in which protein kinase activity can be modulated by reversible oxidation or reduction, involving a unique pair of adjacent cysteines, the “CC” motif. Among all human protein kinases, only LRRK2 contains this “CC” motif (C2024 and C2025) in the Activation Segment (AS) of the kinase domain. In an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, and Gaussian accelerated Molecular Dynamics (GaMD) simulations we could attribute a role for each of those cysteines. We employed reducing and oxidizing agents with potential clinical relevance to investigate effects on kinase activity and microtubule docking. We find that each cysteine gives a distinct contribution: the first cysteine, C2024, is essential for LRRK2 protein kinase activity, while the adjacent cysteine, C2025, contributes significantly to redox sensitivity. Implementing thiolates (R-S-) in GaMD simulations allowed us to analyse how each of the cysteines in the “CC” motif interacts with its surrounding residues depending on its oxidation state. From our studies we conclude that oxidizing agents can downregulate kinase activity of hyperactive LRRK2 PD mutations and may provide promising tools for therapeutic strategies.
Citation
In: npj Parkinson's Disease Volume 10 (2024-04-03) eissn:2373-8057Sponsorship
Gefördert im Rahmen des Projekts DEALCitation
@article{doi:10.17170/kobra-2024082910749,
author={Trilling, Chiara R. and Weng, Jui-Hung and Kaila Sharma, Pallavi and Nolte, Viktoria and Wu, Jian and Ma, Wen and Boassa, Daniela and Taylor, Susan T. and Herberg, Friedrich Wilhelm},
title={RedOx regulation of LRRK2 kinase activity by active site cysteines},
journal={npj Parkinson's Disease},
year={2024}
}
0500 Oax 0501 Text $btxt$2rdacontent 0502 Computermedien $bc$2rdacarrier 1100 2024$n2024 1500 1/eng 2050 ##0##http://hdl.handle.net/123456789/16011 3000 Trilling, Chiara R. 3010 Weng, Jui-Hung 3010 Kaila Sharma, Pallavi 3010 Nolte, Viktoria 3010 Wu, Jian 3010 Ma, Wen 3010 Boassa, Daniela 3010 Taylor, Susan T. 3010 Herberg, Friedrich Wilhelm 4000 RedOx regulation of LRRK2 kinase activity by active site cysteines / Trilling, Chiara R. 4030 4060 Online-Ressource 4085 ##0##=u http://nbn-resolving.de/http://hdl.handle.net/123456789/16011=x R 4204 \$dAufsatz 4170 5550 {{Bioinformatik}} 5550 {{Parkinson-Krankheit}} 5550 {{Proteine}} 7136 ##0##http://hdl.handle.net/123456789/16011
2024-09-02T10:37:52Z 2024-09-02T10:37:52Z 2024-04-03 doi:10.17170/kobra-2024082910749 http://hdl.handle.net/123456789/16011 Gefördert im Rahmen des Projekts DEAL eng Namensnennung 4.0 International http://creativecommons.org/licenses/by/4.0/ computational biology and bioinformatics Parkinson's disease proteins 540 570 RedOx regulation of LRRK2 kinase activity by active site cysteines Aufsatz Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson’s disease (PD). Most of these pathogenic mutations are located in the kinase domain (KD) or GTPase domain of LRRK2. In this study we describe a mechanism in which protein kinase activity can be modulated by reversible oxidation or reduction, involving a unique pair of adjacent cysteines, the “CC” motif. Among all human protein kinases, only LRRK2 contains this “CC” motif (C2024 and C2025) in the Activation Segment (AS) of the kinase domain. In an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, and Gaussian accelerated Molecular Dynamics (GaMD) simulations we could attribute a role for each of those cysteines. We employed reducing and oxidizing agents with potential clinical relevance to investigate effects on kinase activity and microtubule docking. We find that each cysteine gives a distinct contribution: the first cysteine, C2024, is essential for LRRK2 protein kinase activity, while the adjacent cysteine, C2025, contributes significantly to redox sensitivity. Implementing thiolates (R-S-) in GaMD simulations allowed us to analyse how each of the cysteines in the “CC” motif interacts with its surrounding residues depending on its oxidation state. From our studies we conclude that oxidizing agents can downregulate kinase activity of hyperactive LRRK2 PD mutations and may provide promising tools for therapeutic strategies. open access Trilling, Chiara R. Weng, Jui-Hung Kaila Sharma, Pallavi Nolte, Viktoria Wu, Jian Ma, Wen Boassa, Daniela Taylor, Susan T. Herberg, Friedrich Wilhelm doi:10.1038/s41531-024-00683-5 Bioinformatik Parkinson-Krankheit Proteine publishedVersion eissn:2373-8057 npj Parkinson's Disease Volume 10 false 75
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