Aufsatz in einer Fachzeitschrift
Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2
Details zur Publikation
Autor(inn)en: | Schmidt, S.; Weng, J.; Aoto, P.; Boassa, D.; Mathea, S.; Silletti, S.; Hu, J.; Wallbott, M.; Komives, E.; Knapp, S.; Herberg, F.; Taylor, S. |
Publikationsjahr: | 2021 |
Zeitschrift: | Proceedings of the National Academy of Sciences |
Seitenbereich: | e2100844118 |
Jahrgang/Band : | 118 |
Heftnummer: | 23 |
DOI-Link der Erstveröffentlichung: |
Zusammenfassung, Abstract
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 {\textquotedbl}unleashing{\textquotedbl} 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 DYG\textgreeky motif, is the allosteric hub that drives LRRK2 regulation.
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 {\textquotedbl}unleashing{\textquotedbl} 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 DYG\textgreeky motif, is the allosteric hub that drives LRRK2 regulation.
