Structural biology of signaling proteins

RNDr. Veronika Obšilová, Ph.D. — Project head

The Institute of Physiology of the Czech Academy of Sciences

About us

Our group focuses on studying protein-protein interactions with the aim to elucidate the molecular basis of regulatory mechanisms of signaling proteins and their complexes from the biologically important signaling pathways. The main focus is on 14-3-3 protein complexes with proteins involved in the regulation of apoptosis, cancer, G-protein and calcium signaling pathways.

We employ various biophysical (fluorescence spectroscopy, analytical ultracentrifugation, ITC, MST, DSF), structural (X-ray crystallography, NMR, SAXS, HDX-MS) and biochemical (site-directed mutagenesis, enzyme kinetics) approaches to understand the structure-function relationships of studied proteins and protein-protein complexes.

Objectives

Preparation, biochemical and biophysical characterization of selected signaling proteins.
Determination of binding affinities and stoichiometries of studied signaling complexes.
Structural studies of selected signaling protein complexes.

Group Profile (Home Institution): Structural Biology of Signaling Proteins - Institute of Physiology AS CR, v.v.i. (cas.cz)

News

New findings on the structure of the protein kinase ASK1 - a key enzyme in the regulation of the cellular response to stress

Otto Wichterle prize

New Findings On The Structure Of The Foxo4: P53 Complex - A Key Factor In Senescence Regulation

Ph.D. positions available for motivated students

Innovative method of the Forhkead transcription factor FOXO3 regulation

Publications

2024

Obsilova V, Obsil T. (2024) The yeast 14-3-3 proteins Bmh1 and Bmh2 regulate key signaling pathways. Front Mol Biosci. 24:1327014. DOI: 10.3389/fmolb.2024.1327014.

Honzejkova K, Kosek D, Obsilova V, Obsil T. (2024) The cryo-EM structure of ASK1 reveals an asymmetric architecture allosterically modulated by TRX1. Elife 13:RP95199. DOI: 10.7554/eLife.95199.

2023

Masaryk J, Kale D, Pohl P, Ruiz-Castilla FJ, Zimmermannová O, Obšilová V, Ramos J, Sychrová H. (2023) The second intracellular loop of the yeast Trk1 potassium transporter is involved in regulation of activity, and interaction with 14-3-3 proteins. Comput Struct Biotechnol J. 21:2705-2716. DOI: 10.1016/j.csbj.2023.04.019.

Petrvalska O, Honzejkova K, Koupilova N, Herman P, Obsilova V, Obsil T. (2023) 14-3-3 protein inhibits CaMKK1 by blocking the kinase active site with its last two C-terminal helices. Protein Sci. 32:e4805. DOI: 10.1002/pro.4805.

2022

Joshi R, Pohl P, Strachotova D, Herman P, Obsil T, Obsilova V. (2022) Nedd4-2 binding to 14-3-3 modulates the accessibility of its catalytic site and WW domains. Biophys. J. 121(7):1299-1311. DOI: 10.1016/j.bpj.2022.02.025.

Mandal R, Kohoutova K, Petrvalska O, Horvath M, Srb P, Veverka V, Obsilova V and Obsil T (2022) FOXO4 interacts with p53 TAD and CRD and inhibits its binding to DNA. Protein Sci. 31:e4287. DOI: 10.1002/pro.4287

Kohoutova K, Docekal V, Ausserlechner MJ, Kaiser N, Tekel A, Mandal R, Horvath M, Obsilova V, Vesely J, Hagenbuchner J and Obsil T (2022) Lengthening the Guanidine−Aryl Linker of Phenylpyrimidinylguanidines Increases Their Potency as Inhibitors of FOXO3-Induced Gene Transcription. ACS Omega 7(38):34632-34646. DOI: 10.1021/acsomega.2c04613.

Obsilova V and Obsil T (2022) Structural insights into the functional roles of 14-3-3 proteins. Front. Mol. Biosci. 9:1016071. DOI: 10.3389/fmolb.2022.1016071.

2021

Pohl P, Joshi R, Petrvalska O, Obsil T and Obsilova V. (2021) 14-3-3-protein regulates Nedd4-2 by modulating interactions between HECT and WW domains. Commun. Biol. 4(1):899. DOI: 10.1038/s42003-021-02419-0

Horvath M, Petrvalska O, Herman P, Obsilova V and Obsil T. (2021) 14-3-3 proteins inactivate DAPK2 by promoting its dimerization and protecting key regulatory phosphosites. Commun. Biol. 4(1):986. DOI: 10.1038/s42003-021-02518-y

Neves JF, Petrvalska O, Bosica F, Cantrelle FX, Merzougui H, O'Mahony G, Hanoulle X, Obsil T, Landrieu I. (2021) Phosphorylated full-length Tau interacts with 14-3-3 proteins via two short phosphorylated sequences, each occupying a binding groove of 14-3-3 dimer. FEBS J. 288(6):1918-1934. DOI: 10.1111/febs.15574

Bousova K, Bednarova L, Zouharova M, Vetyskova V, Postulkova K, Hofbauerová K, Petrvalska O, Vanek O, Tripsianes K, Vondrasek J. (2021) The order of PDZ3 and TrpCage in fusion chimeras determines their properties-a biophysical characterization. Protein Sci. 30(8):1653-1666. DOI: 10.1002/pro.4107

Obsilova V, Honzejkova K and Obsil T. (2021) Structural Insights Support Targeting ASK1 Kinase for Therapeutic Interventions. Int. J. Mol. Sci. 22(24):13395. DOI:10.3390/ijms222413395.

2020

Kalabova D, Filandr F, Alblova M, Petrvalska O, Horvath M, Man P, Obsil T, Obsilova V. (2020) 14-3-3 protein binding blocks the dimerization interface of caspase-2. FEBS J. 287(16):3494-3510. DOI: 10.1111/febs.15215

Psenakova K, Hexnerova R, Srb P, Obsilova V, Veverka V, Obsil T. (2020) The redox active site of thioredoxin is directly involved in apoptosis signal-regulating kinase 1 binding that is modulated by oxidative stress. FEBS J. 287(8):1626-1644. DOI: 10.1111/febs.15101

Lentini Santo, D., Petrvalska O., Obsilova V., Ottmann C. and Obsil T. (2020) Stabilization of protein-protein interactions between CaMKK2 and 14-3-3 by fusicoccins. ACS Chem Biol. 15(11):3060-3071. DOI: 10.1021/acschembio.0c00821

Obsilova V and Obsil T. (2020) The 14-3-3 Proteins as Important Allosteric Regulators of Protein Kinases. Int. J. Mol. Sci. 21(22):8824. DOI: 10.3390/ijms21228824

Obsilova V and Obsil T. (2020) A new role for 14-3-3 protein in steroidogenesis. FEBS J. 287(18):3921-3924. DOI: 10.1111/febs.15507

2019

Hagenbuchner J, Obsilova V, Kaserer T, Kaiser N, Rass B, Psenakova K, Docekal V, Alblova M, Kohoutova K, Schuster D, Aneichyk T, Vesely J, Obexer P, Obsil T, Ausserlechner MJ. (2019) Modulating FOXO3 transcriptional activity by small, DBD-binding molecules. Elife. 8. pii: e48876. DOI: 10.7554/eLife.48876

Valenti D, Neves JF, Cantrelle FX, Hristeva S, Lentini Santo D, Obsil T, Hanoulle X, Levy LM, Tzalis D, Landrieu I, Ottmann C. (2019) Set-up and screening of a fragment library targeting the 14-3-3 protein interface. Medchemcomm. 10(10):1796-1802. DOI: 10.1039/c9md00215d

Smidova A, Stankova K, Petrvalska O, Lazar J, Sychrova H, Obsil T, Zimmermannova O, Obsilova V. (2019) The activity of Saccharomyces cerevisiae Na⁺, K⁺/H⁺ antiporter Nha1 is negatively regulated by 14-3-3 protein binding at serine 481. Biochim Biophys Acta Mol Cell Res. 1866(12):118534. DOI: 10.1016/j.bbamcr.2019.118534

Psenakova K, Kohoutova K, Obsilova V, Ausserlechner MJ, Veverka V, Obsil T. (2019) Forkhead Domains of FOXO Transcription Factors Differ in both Overall Conformation and Dynamics. Cells. 8(9). pii: E966. DOI: 10.3390/cells8090966

Alblova M, Smidova A, Kalabova D, Lentini Santo D, Obsil T, Obsilova V. (2019) Allosteric activation of yeast enzyme neutral trehalase by calcium and 14-3-3 protein.
Physiol Res. (2019) 68(2):147-160. DOI: 10.33549/physiolres.933950

2018

Smidova A, Alblova M, Kalabova D, Psenakova K, Rosulek M, Herman P, Obsil T, Obsilova V. (2018) 14-3-3 protein masks the nuclear localization sequence of caspase-2. FEBS J. (2018) 285(22):4196-4213. DOI: 10.1111/febs.14670

Kylarova S, Psenakova K, Herman P, Obsilova V, Obsil T. (2018) CaMKK2 kinase domain interacts with the autoinhibitory region through the N-terminal lobe including the RP insert. Biochim. Biophys. Acta-Gen. Subj. 1862(10):2304-2313. DOI: 10.1016/j.bbagen.2018.07.025

Psenakova K, Petrvalska O, Kylarova S, Lentini Santo D, Kalabova D, Herman P, Obsilova V, Obsil T. (2018) 14-3-3 protein directly interacts with the kinase domain of calcium/calmodulin-dependent protein kinase kinase (CaMKK2). Biochim. Biophys. Acta-Gen. Subj. 1862(7):1612-1625. DOI: 10.1016/j.bbagen.2018.04.006

2017

Alblova M, Smidova A, Docekal V, Vesely J, Herman P, Obsilova V, Obsil T. Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1. Proc Natl Acad Sci U S A. 2017, 114:E9811-E9820.

Kalabova D, Smidova A, Petrvalska O, Alblova M, Kosek D, Man P, Obsil T, Obsilova V. Human procaspase-2 phosphorylation at both S139 and S164 is required for 14-3-3 binding. Biochem Biophys Res Commun. 2017, 493:940-945.

Kacirova M, Novacek J, Man P, Obsilova V, Obsil T. Structural Basis for the 14-3-3 Protein-Dependent Inhibition of Phosducin Function. Biophys J. 2017, 112:1339-1349.

2016

Kylarova S, Kosek D, Petrvalska O, Psenakova K, Man P, Vecer J, Herman P, Obsilova V, Obsil T. Cysteine residues mediate high-affinity binding of thioredoxin to ASK1. FEBS J. 2016, 283:3821-3838.

Petrvalska O, Kosek D, Kukacka Z, Tosner Z, Man P, Vecer J, Herman P, Obsilova V, Obsil T. Structural Insight into the 14-3-3 Protein-dependent Inhibition of Protein Kinase ASK1 (Apoptosis Signal-regulating kinase 1). J Biol Chem. 2016, 291:20753-65.

2015

Kacirova M, Kosek D, Kadek A, Man P, Vecer J, Herman P, Obsilova V, Obsil T. Structural Characterization of Phosducin and Its Complex with the 14-3-3 Protein. J Biol Chem. 2015, 290:16246-60.

2014

Kosek D, Kylarova S, Psenakova K, Rezabkova L, Herman P, Vecer J, Obsilova V, Obsil T. Biophysical and structural characterization of the thioredoxin-binding domain of protein kinase ASK1 and its interaction with reduced thioredoxin. J Biol Chem. 2014, 289:24463-74.

Kopecka M, Kosek D, Kukacka Z, Rezabkova L, Man P, Novak P, Obsil T, Obsilova V. Role of the EF-hand-like motif in the 14-3-3 protein-mediated activation of yeast neutral trehalase Nth1. J Biol Chem. 2014, 289:13948-61.

2013

Macakova E, Kopecka M, Kukacka Z, Veisova D, Novak P, Man P, Obsil T, Obsilova V. Structural basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1. Biochim Biophys Acta. 2013, 1830:4491-9.

2012

Veisova D, Macakova E, Rezabkova L, Sulc M, Vacha P, Sychrova H, Obsil T, Obsilova V. Role of individual phosphorylation sites for the 14-3-3-protein-dependent activation of yeast neutral trehalase Nth1. Biochem J. 2012,443:663-670.