The In Situ Structure of Parkinson’s Disease-Linked LRRK2

Watanabe, Reika; Buschauer, Robert; Böhning, Jan; Audagnotto, Martina; Lasker, Keren; Lu, Tsan Wen; Boassa, Daniela; Taylor, Susan S.; Villa, Elizabeth

doi:10.1016/j.cell.2020.08.004

Cited by 167 publications

(258 citation statements)

References 82 publications

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“…Grids with Caulobacter were prepared using cryo-FIB milling as previously described using an Aquilos (Thermo Fisher Scientific) dual-beam SEM equipped 65,66 . Briefly, areas covered with a monolayer of cells were targeted first for coarse milling with an ion beam current of 0.10-0.50 nA, followed by fine milling using 10-50 pA. Lamella width was typically 10-12 um.…”

Section: Methodsmentioning

confidence: 99%

A modular platform for engineering function of natural and synthetic biomolecular condensates

Lasker

Boeynaems

et al. 2021

Preprint

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Phase separation is emerging as a universal principle for how cells use dynamic subcompartmentalization to organize biochemical reactions in time and space. Yet, whether the emergent physical properties of these biomolecular condensates are important for their biological function remains unclear. The intrinsically disordered protein PopZ forms membraneless condensates at the poles of the bacterium Caulobacter crescentus and selectively sequesters kinase-signaling cascades to regulate asymmetric cell division. By dissecting the molecular grammar underlying PopZ phase separation, we find that unlike many eukaryotic examples, where unstructured regions drive condensation, a structured domain of PopZ drives condensation, while conserved repulsive features of the disordered region modulate material properties. By generating rationally designed PopZ mutants, we find that the exact material properties of PopZ condensates directly determine cellular fitness, providing direct evidence for the physiological importance of the emergent properties of biomolecular condensates. Our work codifies a clear set of design principles illuminating how sequence variation in a disordered domain alters the function of a widely conserved bacterial condensate. We used these insights to repurpose PopZ as a modular platform for generating synthetic condensates of tunable function in human cells.

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Section: Methodsmentioning

confidence: 99%

A modular platform for engineering function of natural and synthetic biomolecular condensates

Lasker

Boeynaems

et al. 2021

Preprint

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“…More specifically, the mechanism whereby 14-3-3 protein suppresses LRRK2 kinase activity remains unclear and may involve interference with the dimerization of the ROC domain, modulation of the interaction between ROC and the adjacent COR domain or even interaction of the C-terminal segment with the kinase and COR domains [100,101,103]. Hopefully, the recently reported cryo-TM and cryo-EM structures of LRRK2 will soon be followed by structural studies aimed at clarifying the role of 14-3-3 in LRRK2 function [103,104].…”

Section: Leucine-rich Repeat Protein Kinase-2 (Lrrk2)mentioning

confidence: 99%

The 14-3-3 Proteins as Important Allosteric Regulators of Protein Kinases

Obšilová

Obšil

2020

IJMS

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Phosphorylation by kinases governs many key cellular and extracellular processes, such as transcription, cell cycle progression, differentiation, secretion and apoptosis. Unsurprisingly, tight and precise kinase regulation is a prerequisite for normal cell functioning, whereas kinase dysregulation often leads to disease. Moreover, the functions of many kinases are regulated through protein–protein interactions, which in turn are mediated by phosphorylated motifs and often involve associations with the scaffolding and chaperon protein 14-3-3. Therefore, the aim of this review article is to provide an overview of the state of the art on 14-3-3-mediated kinase regulation, focusing on the most recent mechanistic insights into these important protein–protein interactions and discussing in detail both their structural aspects and functional consequences.

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“…The proposed model indicates the complexity of targeting LRRK2. Villa et al., [21] using cryoEM and integrative modeling, revealed the structure of LRRK2 in situ and showed that the GTPase domain is closer to the microtubule interface, in contrast to the kinase domain which is exposed to the cytoplasm.…”

Section: Figurementioning

confidence: 99%

The tale of proteolysis targeting chimeras (PROTACs) for Leucine‐Rich Repeat Kinase 2 (LRRK2)

et al. 2020

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Here we present the rational design and synthetic methodologies towards proteolysis‐targeting chimeras (PROTACs) for the recently‐emerged target leucine‐rich repeat kinase 2 (LRRK2). Two highly potent, selective, brain‐penetrating kinase inhibitors were selected, and their structure was appropriately modified to assemble a cereblon‐targeting PROTAC. Biological data show strong kinase inhibition and the ability of the synthesized compounds to enter the cells. However, data regarding the degradation of the target protein are inconclusive. The reasons for the inefficient degradation of the target are further discussed.

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The In Situ Structure of Parkinson’s Disease-Linked LRRK2

Cited by 167 publications

References 82 publications

A modular platform for engineering function of natural and synthetic biomolecular condensates

A modular platform for engineering function of natural and synthetic biomolecular condensates

The 14-3-3 Proteins as Important Allosteric Regulators of Protein Kinases

The tale of proteolysis targeting chimeras (PROTACs) for Leucine‐Rich Repeat Kinase 2 (LRRK2)

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