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Environmentally sensitive probes for monitoring protein-membrane interactions at nanomolar concentrations

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F17%3A00475308" target="_blank" >RIV/61388963:_____/17:00475308 - isvavai.cz</a>

  • Result on the web

    <a href="http://dx.doi.org/10.1016/j.bbamem.2017.01.021" target="_blank" >http://dx.doi.org/10.1016/j.bbamem.2017.01.021</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.1016/j.bbamem.2017.01.021" target="_blank" >10.1016/j.bbamem.2017.01.021</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    Environmentally sensitive probes for monitoring protein-membrane interactions at nanomolar concentrations

  • Original language description

    Solvatochromic probes are suitable tools for quantitative characterization of protein-membrane interactions. Based on diverse fluorophores these probes have different fluorescent properties and therefore demonstrate different responses when applied for sensing the interactions of biomolecules. Surprisingly, to the best of our knowledge, no systematic comparison of the sensitivities of solvatochromic dyes for monitoring protein-membrane interactions was described. Hence, a rational choice of an optimal environmentally sensitive probe for such experiments is usually not a straightforward task. In this work we developed a series of thiol-reactive fluorescent probes based on the fluorophores with high sensitivity to their environment and compared them with two widely used DNS and DMN probes. We investigated the responses of these probes to the interaction of probe-labeled presynaptic protein alpha-synuclein with lipid membranes. We observed that newly synthesized probes based on fluorene and chromone dyes, which combine the strongest brightness and significant changes of fluorescence intensity, demonstrated the highest sensitivity to interaction of alpha-synuclein with lipid membranes. They are especially beneficial for sensing in scattering media such as solutions of lipid vesicles. We show that the described probes permit quantitative measurements of ot-synuclein binding to lipid membranes at low nanomolar concentrations. We developed a detailed protocol for measuring Kd and binding stoichiometry for interaction of soluble peripheral proteins with membranes based on the response of the environmentally sensitive fluorescent probes. We applied this protocol for quantification of the affinity of alpha-synuclein to anionic membranes and found that it is substantially higher than it was earlier reported.

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database

  • CEP classification

  • OECD FORD branch

    10608 - Biochemistry and molecular biology

Result continuities

  • Project

  • Continuities

    I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Others

  • Publication year

    2017

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Data specific for result type

  • Name of the periodical

    Biochimica Et Biophysica Acta-Biomembranes

  • ISSN

    0005-2736

  • e-ISSN

  • Volume of the periodical

    1859

  • Issue of the periodical within the volume

    5

  • Country of publishing house

    NL - THE KINGDOM OF THE NETHERLANDS

  • Number of pages

    8

  • Pages from-to

    852-859

  • UT code for WoS article

    000398870800020

  • EID of the result in the Scopus database

    2-s2.0-85012115798