Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2
| Autor: | Steve Silletti, Stefan Knapp, Friedrich W. Herberg, Phillip C. Aoto, Junru Hu, Sven H. Schmidt, Elizabeth A. Komives, Sebastian Mathea, Susan S. Taylor, Maximilian Wallbott, Daniela Boassa, Jui-Hung Weng |
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| Rok vydání: | 2020 |
| Předmět: |
Aging
Allosteric regulation Amino Acid Motifs Hydrogen Deuterium Exchange-Mass Spectrometry GTPase Molecular Dynamics Simulation leucine-rich repeat kinase 2 Neurodegenerative Inhibitory postsynaptic potential Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 Biochemistry Molecular dynamics 03 medical and health sciences 0302 clinical medicine Protein Domains Microtubule Humans 2.1 Biological and endogenous factors Aetiology Kinase activity 030304 developmental biology leucine-rich repeat kinase 2 (LRRK2) 0303 health sciences Multidisciplinary Parkinson's Disease Chemistry Kinase kinase regulation Neurosciences Biological Sciences Gaussian accelerated molecular dynamics LRRK2 Cell biology Brain Disorders nervous system diseases Protein Transport Crosstalk (biology) Protein kinase domain hydrogen-deuterium exchange mass spectrometry (HDX-MS) Docking (molecular) Neurological Parkinson’s disease Biophysics Rab hydrogen-deuterium exchange mass spectrometry 030217 neurology & neurosurgery |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America, vol 118, iss 23 Proceedings of the National Academy of Sciences Proceedings of the National Academy of Sciences of the United States of America |
| DOI: | 10.1101/2020.07.13.198069 |
| Popis: | Significance To achieve a mechanistic understanding of LRRK2, a multidomain protein kinase, we must understand how the conformational landscape is changed by specific mutations that cause LRRK2 to become a driver of Parkinson’s disease (PD). To meet this challenge, we used a construct, LRRK2RCKW, that lacks the N-terminal inhibitory domains. Both catalytic domains as well as full activity are retained in LRRK2RCKW. To capture solvent-exposed/protected regions, we used hydrogen-deuterium exchange mass spectrometry and showed in detail how the conformation changed in the presence of a kinase inhibitor, MLi-2. Using molecular dynamics simulations, we explored the effects of MLi-2 as well as PD mutations on dynamics. Our multitiered analysis defines the kinase domain as a dynamic allosteric hub for LRRK2 activation. To explore how pathogenic mutations of the multidomain leucine-rich repeat kinase 2 (LRRK2) hijack its finely tuned activation process and drive Parkinson’s disease (PD), we used a multitiered approach. Most mutations mimic Rab-mediated activation by “unleashing” kinase activity, and many, like the kinase inhibitor MLi-2, trap LRRK2 onto microtubules. Here we mimic activation by simply deleting the inhibitory N-terminal domains and then characterize conformational changes induced by MLi-2 and PD mutations. After confirming that LRRK2RCKW retains full kinase activity, we used hydrogen-deuterium exchange mass spectrometry to capture breathing dynamics in the presence and absence of MLi-2. Solvent-accessible regions throughout the entire protein are reduced by MLi-2 binding. With molecular dynamics simulations, we created a dynamic portrait of LRRK2RCKW and demonstrate the consequences of kinase domain mutations. Although all domains contribute to regulating kinase activity, the kinase domain, driven by the DYGψ motif, is the allosteric hub that drives LRRK2 regulation. |
| Databáze: | OpenAIRE |
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