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
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Czech description
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Classification
Type
C - Chapter in a specialist book
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
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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
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