RedOx regulation of LRRK2 kinase activity by active site cysteines DOI Creative Commons
Chiara R. Trilling, Jui-Hung Weng, Pallavi Kaila Sharma

и другие.

npj Parkinson s Disease, Год журнала: 2024, Номер 10(1)

Опубликована: Апрель 3, 2024

Abstract Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson’s disease (PD). Most these pathogenic mutations are located in domain (KD) or GTPase LRRK2. In this study we describe a mechanism which protein activity can be modulated by reversible oxidation reduction, involving unique pair adjacent cysteines, “CC” motif. Among all kinases, only LRRK2 contains motif (C2024 C2025) Activation Segment (AS) domain. an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, Gaussian accelerated Molecular Dynamics (GaMD) simulations could attribute role for each those cysteines. We employed reducing oxidizing agents potential clinical relevance to investigate effects on microtubule docking. find that cysteine gives distinct contribution: first cysteine, C2024, is essential activity, while C2025, contributes significantly redox sensitivity. Implementing thiolates (R-S - ) GaMD allowed us analyse how cysteines interacts its surrounding residues depending state. From our studies conclude downregulate hyperactive PD may provide promising tools therapeutic strategies.

Язык: Английский

How are hydrogen peroxide messages relayed to affect cell signalling? DOI Creative Commons
Elizabeth A. Veal, Paraskevi Kritsiligkou

Current Opinion in Chemical Biology, Год журнала: 2024, Номер 81, С. 102496 - 102496

Опубликована: Июль 2, 2024

H

Язык: Английский

Процитировано

8
Oxygen-dependent changes in binding partners and post-translational modifications regulate the abundance and activity of HIF-1α/2α DOI
Leonard A. Daly, Philip Brownridge, Michael Batie

и другие.

Science Signaling, Год журнала: 2021, Номер 14(692)

Опубликована: Июль 20, 2021

The oxygen-dependent regulation of hypoxia-inducible factors is more complex than previously appreciated.

Язык: Английский

Процитировано

34
Classification of Cushing's syndrome PKAc mutants based upon their ability to bind PKI DOI Creative Commons
Mitchell H. Omar, Maryanne Kihiu, Dominic P. Byrne

и другие.

Biochemical Journal, Год журнала: 2023, Номер 480(12), С. 875 - 890

Опубликована: Июнь 12, 2023

Cushing's syndrome is an endocrine disorder caused by excess production of the stress hormone cortisol. Precision medicine strategies have identified single allele mutations within PRKACA gene that drive adrenal syndrome. These promote perturbations in catalytic core protein kinase A (PKAc) impair autoinhibition regulatory subunits and compartmentalization via recruitment into AKAP signaling islands. PKAcL205R found ∼45% patients, whereas PKAcE31V, PKAcW196R, L198insW C199insV insertion mutants are less prevalent. Mass spectrometry, cellular, biochemical data indicate PKAc variants fall two categories: those interact with heat-stable inhibitor PKI, do not. In vitro activity measurements show wild-type W196R activities strongly inhibited PKI (IC50 < 1 nM). contrast, not blocked inhibitor. Immunofluorescent analyses PKI-binding PKAc, E31V, excluded from nucleus protected against proteolytic processing. Thermal stability reveal upon co-incubation metal-bound nucleotide, variant tolerates melting temperatures 10°C higher than PKAcL205. Structural modeling maps PKI-interfering to a ∼20 Å diameter area at active site domain interfaces pseudosubstrate PKI. Thus, kinases individually controlled, compartmentalized, processed through their differential association

Язык: Английский

Процитировано

15
Discovery of a Cushing’s syndrome protein kinase A mutant that biases signaling through type I AKAPs DOI Creative Commons
Mitchell H. Omar, Dominic P. Byrne, Safal Shrestha

и другие.

Science Advances, Год журнала: 2024, Номер 10(8)

Опубликована: Фев. 21, 2024

Adrenal Cushing's syndrome is a disease of cortisol hypersecretion often caused by mutations in protein kinase A catalytic subunit (PKAc). Using personalized medicine screening platform, we discovered driver mutation, PKAc-W196G, ~20% patient samples analyzed. Proximity proteomics and photokinetic imaging reveal that PKAc

Язык: Английский

Процитировано

6
Post-Translational Modifications of the Energy Guardian AMP-Activated Protein Kinase DOI Open Access
Ashley J. Ovens, John W. Scott, Christopher G. Langendorf

и другие.

International Journal of Molecular Sciences, Год журнала: 2021, Номер 22(3), С. 1229 - 1229

Опубликована: Янв. 27, 2021

Physical exercise elicits physiological metabolic perturbations such as energetic and oxidative stress; however, a diverse range of cellular processes are stimulated in response to combat these challenges maintain energy homeostasis. AMP-activated protein kinase (AMPK) is highly conserved enzyme that acts fuel sensor central this adaptive exercise. The complexity AMPK’s role modulating signalling cascades well documented, yet aside from its well-characterised regulation by activation loop phosphorylation, AMPK further subject multitude additional regulatory stimuli. Therefore, review we comprehensively outline current knowledge around the post-translational modifications AMPK, including novel phosphorylation sites, underappreciated roles for ubiquitination, sumoylation, acetylation, methylation oxidation. We provide insight into ramifications modifications, which not only affect activity, but also subcellular localisation, nutrient interactions stability. Lastly, highlight gaps area research perspectives on how field can apply greater rigour characterisation modifications.

Язык: Английский

Процитировано

29
Aurora A kinase activation: Different means to different ends DOI Creative Commons
Nicolas Tavernier, Frank Sicheri, Lionel Pintard

и другие.

The Journal of Cell Biology, Год журнала: 2021, Номер 220(9)

Опубликована: Июль 9, 2021

Aurora A is a serine/threonine kinase essential for mitotic entry and spindle assembly. Recent molecular studies have revealed the existence of multiple, distinct mechanisms activation, each occurring at specific subcellular locations, optimized cellular context, primed by signaling events including phosphorylation oxidation.

Язык: Английский

Процитировано

29
BCAT1 redox function maintains mitotic fidelity DOI Creative Commons

Liliana Francois,

Pavle Boskovic, Julian Knerr

и другие.

Cell Reports, Год журнала: 2022, Номер 41(3), С. 111524 - 111524

Опубликована: Окт. 1, 2022

The metabolic enzyme branched-chain amino acid transaminase 1 (BCAT1) drives cell proliferation in aggressive cancers such as glioblastoma. Here, we show that BCAT1 localizes to mitotic structures and has a non-metabolic function regulator. Furthermore, is required for chromosome segregation cancer induced pluripotent stem cells tumor growth human cerebral organoid mouse syngraft models. Applying gene knockout rescue strategies, the CXXC redox motif crucial controlling cysteine sulfenylation specifically cells, promoting Aurora kinase B localization centromeres, securing accurate segregation. These findings offer an explanation well-established role of proliferation. In summary, our data establish component apparatus safeguards fidelity through moonlighting functionality.

Язык: Английский

Процитировано

22
Structural basis for CEP192-mediated regulation of centrosomal AURKA DOI Creative Commons

Jin-Gyeong Park,

Hanul Jeon, Sang Chul Shin

и другие.

Science Advances, Год журнала: 2023, Номер 9(16)

Опубликована: Апрель 21, 2023

Aurora kinase A (AURKA) performs critical functions in mitosis. Thus, the activity and subcellular localization of AURKA are tightly regulated depend on diverse factors including interactions with multiple binding cofactors. How these different cofactors regulate to elicit levels at distinct locations times is poorly understood. Here, we identified a conserved region CEP192, major cofactor AURKA, that mediates interaction AURKA. Quantitative studies were performed map helix (Helix-1) within CEP192. The crystal structure Helix-1 bound revealed site from other proteins such as TPX2. Inhibiting between cells led mitotic defects, demonstrating importance interaction. Collectively, structural basis for CEP192-mediated regulation centrosome, which TPX2-mediated spindle microtubule.

Язык: Английский

Процитировано

13
Custom Workflow for the Confident Identification of Sulfotyrosine-Containing Peptides and Their Discrimination from Phosphopeptides DOI Creative Commons
Leonard A. Daly, Dominic P. Byrne, Simon Perkins

и другие.

Journal of Proteome Research, Год журнала: 2023, Номер 22(12), С. 3754 - 3772

Опубликована: Ноя. 8, 2023

Protein tyrosine sulfation (sY) is a post-translational modification (PTM) catalyzed by Golgi-resident tyrosyl protein sulfo transferases (TPSTs). Information on sY in humans currently limited to ∼50 proteins, with only handful having verified sites of sulfation. As such, the contribution regulation biological processes remains poorly defined. Mass spectrometry (MS)-based proteomics method choice for PTM analysis but has yet be applied systematic investigation "sulfome", primarily due issues associated discrimination sY-containing from phosphotyrosine (pY)-containing peptides. In this study, we developed an MS-based workflow sY-peptide characterization, incorporating optimized Zr4+ immobilized metal-ion affinity chromatography (IMAC) and TiO2 enrichment strategies. Extensive characterization panel sY- pY-peptides using array fragmentation regimes (CID, HCD, EThcD, ETciD, UVPD) highlighted differences generation site-determining product ions allowed us develop strategy differentiating sulfated peptides nominally isobaric phosphopeptides based low collision energy-induced neutral loss. Application our "sulfomics" HEK-293 cell extracellular secretome facilitated identification 21 new sulfotyrosine-containing several which validate enzymatically, reveals interplay between enzymes relevant both glycan

Язык: Английский

Процитировано

13
Dimeric Structure of the Pseudokinase IRAK3 Suggests an Allosteric Mechanism for Negative Regulation DOI Creative Commons
Sven M. Lange,

Marina I. Nelen,

Philip Cohen

и другие.

Structure, Год журнала: 2020, Номер 29(3), С. 238 - 251.e4

Опубликована: Ноя. 24, 2020

Interleukin-1 receptor associated kinases (IRAKs) are key players in innate immune signaling that mediate the host response to pathogens. In contrast active IRAK1 and IRAK4, IRAK2 IRAK3 pseudokinases lacking catalytic activity their functions poorly understood. is thought be a negative regulator of mutations with asthma cancer. Here, we report crystal structure human pseudokinase domain closed, pseudoactive conformation. dimerizes unique way through head-to-head arrangement not observed any other kinases. Multiple conserved cysteine residues imply potential redox control conformation dimerization. By analyzing asthma-associated mutations, identify an evolutionarily surface on could form interaction interface suggesting model for regulation IRAK4 by IRAK3.

Язык: Английский

Процитировано

31