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Title of the item:

Application of Two-Dimensional Difference Gel Electrophoresis in Identification of Factors Responsible for Virulence of Staphylococcus aureus.

Title :
Application of Two-Dimensional Difference Gel Electrophoresis in Identification of Factors Responsible for Virulence of Staphylococcus aureus.
Authors :
Bonar E; Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Chlebicka K; Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Dubin G; Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.; Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Wladyka B; Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland. .
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Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2020; Vol. 2069, pp. 139-154.
Publication Type :
Journal Article; Research Support, Non-U.S. Gov't
Language :
English
Imprint Name(s) :
Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,
MeSH Terms :
Electrophoresis, Gel, Two-Dimensional*
Proteomics*
Staphylococcal Infections/*metabolism
Staphylococcus aureus/*metabolism
Staphylococcus aureus/*pathogenicity
Virulence Factors/*biosynthesis
Animals ; Humans ; Two-Dimensional Difference Gel Electrophoresis
Contributed Indexing :
Keywords: 2D DIGE*; Electrophoresis*; Protein*; Proteome*; Proteomics*; Staphylococcus aureus*; Virulence*
Substance Nomenclature :
0 (Virulence Factors)
Entry Date(s) :
Date Created: 20190917 Date Completed: 20201217 Latest Revision: 20201217
Update Code :
20210623
DOI :
10.1007/978-1-4939-9849-4_11
PMID :
31523772
Academic Journal
Staphylococcus aureus is a dangerous opportunistic pathogen of humans and animals. Highly virulent and multi-antibiotic-resistant strains are of particular concern due to high invasiveness and limited array of useful treatment options. Proteomics allows identification and investigation of staphylococcal virulence factors to better understand and treat the related disease. Two-dimensional difference gel electrophoresis (2D DIGE) is a powerful method for identification of differences in staphylococcal proteomes, both intracellular and secretory. Not only the presence of particular proteins and their quantities may be determined, but also each modification changing the molecular mass and/or isoelectric point of a protein is trackable. Especially, 2D DIGE allows for detection of posttranslational modifications, including processing and degradation by proteases. For differential analysis, protein samples are labeled with spectrally distinguishable fluorescent dyes, mixed and separated according to their isoelectric point (first dimension), and then electrophoresed in the presence of sodium dodecyl sulfate according to their molecular mass (second dimension). Exceptional resolution of 2D DIGE allows to obtain focused and sharp protein spots, and identify a large number of differentiating proteins. Here we provide protocols for TRI Reagent-based preparation of high-quality samples for 2D DIGE, sample separation, and ways of handling differentiating protein spots which lead to samples ready for protein identification using MS.

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