Finished grants:

  • Role of acute myeloid leukemia-associated nucleophosmin mutations in the NPM-p53-Mdm2 regulatory network

    Grant agency: GACR
    Identification number of grant: 22-03875S
    Principal investigator: prof. RNDr. Petr Heřman, CSc.
    Co-Principal investigators: Mgr. Barbora Brodská, Ph.D.

    Grant annotation

  • FluoTRAM: Fluorescence-detected Transient Absorption Microscopy

    Grant agency: EU-MSCA
    Identification number of grant: 102-56/21546
    Principal investigator: RNDr. Pavel Malý, Ph.D.

    Grant annotation

    Fluorescence microscopy is an indispensable tool in many areas of research. In life sciences it has been perfected for biological sample imaging either by its autofluorescence or using fluorescent markers such as dyes or fluorescent proteins. It is thus possible to localize molecules in cells, obtaining information on their dynamics and environment. The fluorescence detection, by its nature, provides information only about on the final, emissive state of the molecules after photoexcitation. Meanwhile, transient absorption spectroscopy enables to track the initial state of the molecules after absorption and the following excitation dynamics. However, such ultrafast nonlinear techniques typically require volume samples and are detected coherently. We have recently developed a new way to measure transient absorption by detecting the sample fluorescence. In project FluoTRAM, we will implement our technique in the fluorescence microscope, where it truly reveals its potential. Using the established imaging techniques and markers, FluoTRAM brings the additional information on the excitation event and the dynamics towards the emissive state. This comprehensive additional information will be of great use in life sciences and beyond, examples include correlation of the excitation and emission spectra (increased Stokes shift vs red shift) for dye probes, intramolecular charge transfer in fluorescent proteins, or charge transfer and recombination in organic materials.

    For details see the webpage of the project .

    EU MSCA grant number 101030656

  • Role of nucleophosmin interactome in acute myeloid leukemia with mutated NPM

    Grant agency: GACR
    Identification number of grant: 19-04099S
    Principal investigator: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    One third of acute myeloid leukemia (AML) occurrences exhibits characteristic nucleophosmin
    (NPM) mutation causing its aberrant cytoplasmic localization. Subsequently, multiple NPMinteraction
    partners, interacting also with mutated NPM or its complexes, become mislocalized
    from their site-of-action with functional and regulatory consequences. By combination of
    spectroscopic, microscopic, biochemical and immunochemical methods (e.g. FLIM, FRET, GFPTrap)
    we will evaluate unknown/neglected relations between AML-related NPM mutations,
    oligomerization, and ability of wild-type and mutated NPM to interact with the tumor suppressor
    p53. Dynamics and cellular trafficking of NPM and p53 will be assessed in vivo and the role of
    NPM-interacting proteins regulated by p53 in the leukemogenesis will be elucidated. We will
    characterize cellular response of the NPM interaction network to anticancer drug treatment in
    relation to the NPM mutation and the drug used. Results will contribute to understanding of the
    role of NPM mutation in leukemogenesis and will open new strategies for AML treatment.

  • Nanobiophotonics for future health care

    Grant agency: GB
    Identification number of grant: CE P205/12/G118
    Principal investigator: Doc. Ing. Jiří Homola, CSc., DSc.
    Co-Principal investigators: prof. RNDr. Josef Štěpánek, CSc.

    Grant annotation

    As modern medicine evolves towards quantitative and molecular based science, biophotonics is envisioned to play an increasingly important role in multiple areas of medicine, contributing to quality of health care, reduction of health care costs, and sustainability of the medical care in the ageing society. The proposal aims to advance research in selected areas of nanobiophotonics with focus on photonic molecular biosensors based on plasmonic nanostructures. The main areas of research in this project include research into plasmonic phenomena on metallic nanostructures, development of novel tools for analysis and design of plasmonic nanostructures, fabrication and experimental characterization of plasmonic nanostructures with potential for surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS) sensing, interfacing biomolecules with inorganic nanostructures and investigation of interactions between such biophotonic structures and biological samples, and realization of SPR and SERS biosensors for detection of biomarkers of onco-hematological diseases.

  • Role of the 14-3-3 protein in the regulation of phosducin function

    Grant agency: GACR
    Identification number of grant: GACR P305/11/0708
    Principal investigator: Doc. RNDr. Tomáš Obšil, Ph.D.

    Grant annotation

    The main goal is to study the structural basis for the 14-3-3 protein-dependent regulation of phosducin function. Methods of fluorescence spectroscopy, analytical ultracentrifugation and protein crystallography will be used as principal tools. Phosducin specifically binds with high affinity to Gtβg heterodimer and inhibits its interaction with Gtα or other effectors. Thus phosducin down-regulates the light response in photoreceptors. Phosducin function is regulated through the phosphorylation at Ser54 and Ser73 and the binding to the regulatory 14-3-3 protein. The detailed mechanism of the 14-3-3 protein-dependent inhibition of phosducin function is unknown. We propose to perform structural studies of the 14-3-3/phosducin complex, to identify phosducin regions involved in the interaction with 14-3-3 and thus to mechanistically explain the inhibitory role of 14-3-3 in phosducin regulation. The proposed research will increase our understanding concerning not only the role of the 14-3-3 protein in phosducin regulation (and G protein signaling) but the 14-3-3 protein function in general as well.

  • Role of membrane potential in the lateral microdomain organization of the plasma membrane

    Grant agency: GACR
    Identification number of grant: GACR P205/12/0720
    Principal investigator: prof. RNDr. Petr Heřman, CSc.
    Co-Principal investigators: RNDr. Jan Malínský, Ph.D.

    Grant annotation

    epending on their actual chemical composition, membranes are subdivided into dynamic microdomains differing in size, lipid order and physical properties. Rearrangement of the microdomains strongly modulates biochemical processes on the membrane and cellular responses. The main goal of the project is to determine, how changes of the membrane potential influence lateral reconfiguration of the plasma membrane microdomains and, consequently, physical properties of the membrane. We will study relation between potential-induced domain reconfiguration and susceptibility of cells to stressors. Dynamics of the process and role of particular lipids and proteins will be studied as well. Fluorescence spectroscopy and biochemical methods will be used as main tools. Results should shed light on the role of membrane potential and microdomain structure in the process of the first cellular response to changed external conditions. With respect to the common principles of membrane organization through the phylogeny, a general biological impact is anticipated.

  • Study of the interaction between 14-3-3 protein and regulator of G protein signaling RGS3

    Grant agency: GAAV
    Identification number of grant: IAA501110801
    Principal investigator: Doc. RNDr. Tomáš Obšil, Ph.D.
    Co-Principal investigators: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    The main goal of this project is to understand the structural basis of 14-3-3 protein–dependent regulation of function of RGS3 protein. 14-3-3 proteins are regulatory molecules that bind to other proteins in a phosphorylation dependent manner. RGS proteins function both as GTPase activating proteins for G proteins and regulators of G protein-effector interactions. RGS3 protein is one of the RGS proteins known to be 14-3-3 binding partners. The role of 14-3-3 protein in the regulation of RGS3 seems to be an inhibition of its GTPase activating function on G protein mediated signals. However structural details concerning interaction between 14-3-3 and RGS3 are unknown. We propose to perform (1) biophysical characterization of 14-3-3/RGS3 complex; (2) to study how 14-3-3 protein inhibits the interaction between RGS3 protein and Galpha subunit of G protein using fluorescence spectroscopy; (3) to perform structural analysis of the 14-3-3/RGS3 complex using protein X-ray crystallography.

  • Self-organization principles of non-membrane-bound organels in eukaryotic cells

    Grant agency: GACR
    Identification number of grant: GACR 204/07/0133
    Principal investigator: RNDr. Jan Malínský, Ph.D.

    Grant annotation

    The non-membrane-bound structures represent a unique problem in the cell biology. They are stable in space and time although they continuously exchange their components with the surrounding environment. In this project we propose to test a hypothesis that these structures behave as objects in a steady-state-balance, which can be described by self-organization principles. The strategy of this proposal is to acquire experimental data about localization, dynamics and interactions of individual proteins in different cellular organelles and, based on these data, model formation and maintenance of these structures. We aim to apply self-organization models on three cellular non-membrane bound compartments: the nucleolus, the Cajal body and RMC C (Raft-based Membrane Compartment C or eisosomes) and one nuclear process – DNA replication and test the acquired models experimentally. We believe that the models will open new directions in the research of cellular structures and their functions.

  • Biophysical study of MntH membrane transport proteins: structure-function relationship

    Grant agency: GACR
    Identification number of grant: GACR 204/07/0558
    Principal investigator: RNDr. Roman Chaloupka, Ph.D.

    Grant annotation

    This project aims at bacterial transporters of divalent metal ions (DMI) from the Nramp/MntH family counting among the proton-dependent secondary active membrane transport proteins. Although the Nramp-related proteins, playing a key role in various physiological processes including macrophage-pathogen interaction, are present in virtually all organisms, the transport mechanism is not understood. We propose to advance knowledge of the structure-function relationship in the MntH/Nramp family by studying the function of key residues in vivo, in vitro (right-side out vesicles, in-side out vesicles and proteoliposomes) and in a peptide-membrane model system. The peptide model allows complementing the functional assays by information about transmembrane segment structure, insertion and folding, and possibly some functional details.

  • Role of the phosphorylation and the 14-3-3 protein binding in the regulation of signaling complexes function

    Grant agency: GACR
    Identification number of grant: GACR 204/06/0565
    Principal investigator: Doc. RNDr. Tomáš Obšil, Ph.D.
    Co-Principal investigators: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    The overall goal of this project is to understand the structural basis of 14-3-3 protein-dependent regulation of function of two biologically important signaling complexes: 14-3-3 protein/forkhead transcription factor FoxO4 and 14-3-3 protein/tyrosine hydroxylase complex. 14-3-3 proteins are regulatory molecules that bind to other proteins in a phosphorylation dependent manner. Forkhead transcription factor FoxO4 is involved in the metabolism control, apoptosis or oncogenesis, and its activity is controlled through the phosphorylation and the 14-3-3 protein binding. Tyrosine hydroxylase is an enzyme responsible for catalysis of the first step in the biosynthesis of catecholamines and its activity is also regulated through the phosphorylation and the 14-3-3 protein binding. In both cases the molecular mechanism of the 14-3-3 protein-dependent regulation of target protein function is unknown.

  • Role of adaptation in structure-function relations of microorganisms

    Grant agency: NATO
    Identification number of grant: NATO-CNR 218.2049
    Principal investigator: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    not available

  • Fluorescence microscopy with time and spectral resolution

    Grant agency: GAUK
    Identification number of grant: GAUK 171/2003
    Principal investigator: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    not available

  • Role of selected cellular components on kinetics and redistribution of potential-sensitive fluorescent probes

    Grant agency: GAUK
    Identification number of grant: GAUK 168/1996
    Principal investigator: prof. RNDr. Petr Heřman, CSc.

    Grant annotation

    not available