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Identification of Protein Interaction Partners in Bacteria Using Affinity Purification and SILAC Quantitative Proteomics

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60162694%3AG44__%2F24%3A00558811" target="_blank" >RIV/60162694:G44__/24:00558811 - isvavai.cz</a>

  • Result on the web

    <a href="https://link.springer.com/book/10.1007/978-1-0716-2863-8" target="_blank" >https://link.springer.com/book/10.1007/978-1-0716-2863-8</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.1007/978-1-0716-2863-8_3" target="_blank" >10.1007/978-1-0716-2863-8_3</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    Identification of Protein Interaction Partners in Bacteria Using Affinity Purification and SILAC Quantitative Proteomics

  • Original language description

    Affinity purification, combined with mass spectrometry (AP-MS) is considered a pivotal technique in protein–protein interaction studies enabling systematic detection at near physiological conditions. The addition of a quantitative proteomic method, like SILAC metabolic labeling, allows the elimination of non-specifically bound contaminants which greatly increases the confidence of the identified interaction partners. Compared to eukaryotic cells, the SILAC labeling of bacteria has specificities that must be considered. The protocol presented here describes the labeling of bacterial cultures with stable isotope-labeled amino acids, purification of an affinity-tagged protein, and sample preparation for MS measurement. Finally, we discuss the analysis and interpretation of MS data to identify and select the specific partners interacting with the protein of interest. As an example, this workflow is applied to the discovery of potential interaction partners of glyceraldehyde-3-phosphate dehydrogenase in the gram-negative bacterium Francisella tularensis.

  • Czech name

  • Czech description

Classification

  • Type

    C - Chapter in a specialist book

  • 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

    2023

  • Confidentiality

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

Data specific for result type

  • Book/collection name

    SILAC. Methods and Protocols, vol. 2603

  • ISBN

    978-1-0716-2862-1

  • Number of pages of the result

    12

  • Pages from-to

    31-42

  • Number of pages of the book

    286

  • Publisher name

    Humana

  • Place of publication

    New York

  • UT code for WoS chapter