A5087618958- Ubiquitin and proteasome pathways
- Protein Degradation and Inhibitors
- interferon and immune responses
- PI3K/AKT/mTOR signaling in cancer
- NF-κB Signaling Pathways
- Immune Response and Inflammation
- Peptidase Inhibition and Analysis
- Metabolism, Diabetes, and Cancer
- Cancer-related gene regulation
- Protein Kinase Regulation and GTPase Signaling
- Wnt/β-catenin signaling in development and cancer
- Cytokine Signaling Pathways and Interactions
- Cellular transport and secretion
- Pancreatic function and diabetes
- Glycosylation and Glycoproteins Research
- Histone Deacetylase Inhibitors Research
- Retinal Development and Disorders
- Cancer-related Molecular Pathways
- Epigenetics and DNA Methylation
- Microtubule and mitosis dynamics
- CRISPR and Genetic Engineering
- Melanoma and MAPK Pathways
- Signaling Pathways in Disease
- Nerve injury and regeneration
- TGF-β signaling in diseases
MRC Protein Phosphorylation and Ubiquitylation Unit
2012-2024
University of Dundee
2011-2024
Medical Research Council
2014-2024
The specificities of 65 compounds reported to be relatively specific inhibitors protein kinases have been profiled against a panel 70–80 kinases. On the basis this information, effects that we studied in cells and other data literature, recommend use following small-molecule inhibitors: SB 203580/SB202190 BIRB 0796 used parallel assess physiological roles p38 MAPK (mitogen-activated kinase) isoforms, PI-103 wortmannin inhibit phosphatidylinositol (phosphoinositide) 3-kinases, PP1 or PP2 with...
We have studied the mechanism of A-769662, a new activator AMP-activated protein kinase (AMPK). Unlike other pharmacological activators, it directly activates native rat AMPK by mimicking both effects AMP, i.e. allosteric activation and inhibition dephosphorylation. found that has no effect on isolated α subunit domain, with or without associated autoinhibitory interaction glycogen β glycogen-binding domain. Although mimics actions binding AMP to Bateman domains γ subunit. The addition...
The LKB1 tumour suppressor phosphorylates and activates AMPK (AMP-activated protein kinase) when cellular energy levels are low, thereby suppressing growth through multiple pathways, including inhibiting the mTORC1 (mammalian target of rapamycin complex 1) kinase that is activated in majority human cancers. Blood glucose-lowering Type 2 diabetes drugs also induce to activate AMPK, indicating these compounds could be used suppress cells. In present study, we investigated importance LKB1–AMPK...
The Vps34 (vacuolar protein sorting 34) class III PI3K (phosphoinositide 3-kinase) phosphorylates PtdIns (phosphatidylinositol) at endosomal membranes to generate PtdIns(3)P that regulates membrane trafficking processes via its ability recruit a subset of proteins possessing PtdIns(3)P-binding PX (phox homology) and FYVE domains. In the present study, we describe highly selective potent inhibitor Vps34, termed VPS34-IN1, inhibits with 25 nM IC50 in vitro, but does not significantly inhibit...
Leucine-rich repeat kinase 2 (LRRK2) is one of the most promising targets for Parkinson's disease. LRRK2-targeting strategies have primarily focused on type 1 inhibitors, which, however, limitations as inhibited protein can interfere with natural mechanisms, which could lead to undesirable side effects. Herein, we report development LRRK2 proteolysis targeting chimeras (PROTACs), culminating in discovery degrader XL01126, an alternative strategy. Initial designs and screens PROTACs based...
We detected a protein in rabbit skeletal muscle extracts that was phosphorylated rapidly by SGK1 (serum- and glucocorticoid-induced kinase 1), but not Bα, identified it as NDRG2 (N-myc downstream-regulated gene 2). at Thr330, Ser332 Thr348in vitro. All three residues were from wild-type mice, mice do express SGK1. also the related NDRG1 isoform Thr328, Ser330 Thr346 (equivalent to Thr348 of NDRG2), well Thr356 Thr366. Residues Thr346, Thr366 are located within identical decapeptide sequences...
The compound BIRB796 inhibits the stress-activated protein kinases p38alpha and p38beta is undergoing clinical trials for treatment of inflammatory diseases. Here we report that also activity activation SAPK3/p38gamma. This occurs at higher concentrations than those inhibit lower JNK isoforms. We show these concentrations, blocks stress-induced phosphorylation scaffold SAP97, further establishing this a physiological substrate Our results demonstrate BIRB796, in combination with SB203580,...
The compound BAY 11-7082 inhibits IκBα [inhibitor of NF-κB (nuclear factor κB)α] phosphorylation in cells and has been used to implicate the canonical IKKs (IκB kinases) >350 publications. In present study we report that does not inhibit IKKs, but suppresses their activation LPS (lipopolysaccharide)-stimulated RAW macrophages IL (interleukin)-1-stimulated IL-1R (IL-1 receptor) HEK (human embryonic kidney)-293 cells. exerts these effects by inactivating E2-conjugating enzymes Ubc...
Autosomal dominant mutations that activate the leucine-rich repeat kinase 2 (LRRK2) cause inherited Parkinson's disease. Recent work has revealed LRRK2 directly phosphorylates a conserved threonine/serine residue in effector-binding switch-II motif of number Rab GTPase proteins, including Rab10. Here we describe facile and robust method to assess phosphorylation endogenous Rab10 mouse embryonic fibroblasts (MEFs), lung spleen-derived B-cells, based on ability Phos-tag reagent retard...
AMPK is a sensor of cellular energy status and promising target for drugs aimed at metabolic disorders. We have studied the selectivity mechanism recently described activator, C2, its cell-permeable prodrug, C13. C2 was potent allosteric activator α1-complexes that, like AMP, also protected against Thr172 dephosphorylation. Compared with caused only partial activation α2-complexes failed to protect them show that both effects could be fully restored by exchanging part linker between...
We have reported previously that activation of the MyD88-signaling network rapidly induces formation hybrid ubiquitin chains containing both Lys63-linked and Met1-linked (Ub) oligomers, some which are attached covalently to Interleukin Receptor Associated kinase 1. Here we show Lys63/Met1-Ub hybrids also formed when TNFR1/TRADD, TLR3/TRIF- NOD1/RIP2-signaling networks activated, Receptor-Interacting Protein 1 (TNFR1 pathway) or 2 (NOD1 pathway). These observations suggest general...
SGK3 is a PX domain containing protein kinase activated at endosomes downstream of class 1 and 3 PI3K family members by growth factors oncogenic mutations. plays key role in mediating resistance breast cancer cells to or Akt inhibitors, substituting for the loss activity restoring proliferative pathways such as mTORC1 signaling. It therefore critical develop tools potently target obstruct its inhibitor resistance. Here, we describe development SGK3-PROTAC1, PROTAC conjugate 308-R SGK with...
Proteolysis-targeting chimeras (PROTACs) bring a protein of interest (POI) into spatial proximity an E3 ubiquitin ligase, promoting POI ubiquitylation and proteasomal degradation. PROTACs rely on endogenous cellular machinery to mediate degradation, therefore the subcellular location access ligase being recruited potentially impacts PROTAC efficacy. To interrogate whether context influences PROTAC-mediated we expressed either Halo or FKBP12F36V (dTAG) constructs consisting varying...
Many phosphorylation signal transduction pathways in the eukaryotic cell are modulated by posttranslational modification of specific serines/threonines with N-acetylglucosamine (O-GlcNAc). Levels O-GlcNAc on key proteins regulate biological processes as diverse cycle, insulin signaling, and protein degradation. The two enzymes involved this dynamic abundant transferase O-GlcNAcase. Structural data have recently revealed that O-GlcNAcase possesses an active site significant structural...
AbstractVX-680, also known as MK-0457, is a member of diverse group small molecules that inhibit the Aurora kinases, and has shown significant potential an anti-cancer agent. In keeping with many protein kinase inhibitors, this compound not monospecific agent, its cellular specificity remains largely unknown. cells, VX-680 blocks mitotic Histone H3 phosphorylation induces polyploidy apoptosis, consistent inhibition B. study, we have investigated effects in proliferating human cancer...
Ubiquitin and UBL (ubiquitin-like) modifiers are small proteins that covalently modify other to alter their properties or behaviours. modification (ubiquitylation) targets many substrates, often leading proteasomal degradation. NEDD8 (neural-precursor-cell-expressed developmentally down-regulated 8) is the most closely related ubiquitin, its best-studied role activation of CRLs (cullin-RING ubiquitin ligases) by conjugation a conserved C-terminal lysine residue on cullin proteins. The...
Significance NF-κB and IFN Regulatory Factor 5 (IRF5) are required for the transcription of many proinflammatory cytokines in myeloid cells. The protein kinase IKKβ is major activator but how IRF5 activated has been unclear. This paper demonstrates that also activates by catalyzing phosphorylation Ser462. this serine induces dimerization its translocation to nucleus. activation master factors innate immune system same provides a mechanism coordinated control response inflammatory stimuli.
Article1 August 2016Open Access Source DataTransparent process The hVps34-SGK3 pathway alleviates sustained PI3K/Akt inhibition by stimulating mTORC1 and tumour growth Ruzica Bago Corresponding Author MRC Protein Phosphorylation Ubiquitylation Unit, College of Life Sciences, University Dundee, UK Search for more papers this author Eeva Sommer Pau Castel Human Oncology Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA Claire Crafter iMED, AstraZeneca, CRUK...
Mutations leading to inappropriate activation of Akt isoforms contribute proliferation and survival a significant proportion human cancers. is activated by phosphorylation its T-loop residue (Thr308) PDK1 (3-phosphoinositide-dependent kinase-1) C-terminal hydrophobic motif (Ser473) mTORC2 [mTOR (mammalian target rapamycin) complex 2]. Potent inhibitors such as GSK2334470 have recently been elaborated potential anti-cancer agents. However, these compounds were surprisingly ineffective at...
The LKB1 tumour suppressor protein kinase functions to activate two isoforms of AMPK (AMP-activated kinase) and 12 members the AMPK-related family kinases. highly conserved C-terminal residues are phosphorylated (Ser431) by PKA (cAMP-dependent RSK (ribosomal S6 farnesylated (Cys433) within a CAAX motif. To better define role that these post-translational modifications play, we created homozygous LKB1S431A/S431A LKB1C433S/C433S knockin mice. These animals were viable, fertile displayed no...
Plasmacytoid dendritic cells (pDCs) are characterized by their ability to produce high levels of type 1 interferons in response ligands that activate TLR7 and TLR9, but the signaling pathways required for IFN production incompletely understood. Here we exploit human pDC cell line Gen2.2 improved pharmacological inhibitors protein kinases address this issue. We demonstrate TLR9 require TAK1-IKKβ pathway induce IFNβ via a is independent degradation IκBα. also show IKKβ activity, as well...
Research Article16 April 2018Open Access Source DataTransparent process Myeloid cell deficiency of p38γ/p38δ protects against candidiasis and regulates antifungal immunity Dayanira Alsina-Beauchamp Department Immunology Oncology, Centro Nacional de Biotecnología/CSIC, Madrid, Spain Search for more papers by this author Alejandra Escós orcid.org/0000-0002-2990-7920 Pilar Fajardo Diego González-Romero Ester Díaz-Mora Ana Risco Miguel A Martín-Serrano Carlos del Fresno Immunobiology...