Proteomics

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Improved SILAC Method for Double Labeling of Bacterial Proteome


ABSTRACT: Stable isotope labeling with amino acids in cell culture (SILAC) is a robust proteomics method with the advantages of reproducibility and easy handling. This method is popular for the analysis of mammalian cells. However, amino acid conversion in bacteria decreases the labeling efficiency and quantification accuracy, limiting the application of SILAC in bacterial proteomics to auxotrophic bacteria or single labeling with lysine. In this study, we found that adding high concentrations of isotope-labeled (heavy) and natural (light) amino acids into SILAC minimal medium can efficiently inhibit the complicated amino acid conversions between each other. This simple and straightforward strategy facilitated the full incorporation of amino acids into the bacterial proteome with good accuracy. The high labeling efficiency can be reached in different bacteria by slightly modifying the supplement of amino acids in culture media, promoting the widespread application of SILAC technique in bacterial proteomics.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Escherichia Coli Staphylococcus Aureus

SUBMITTER: zhao xinlu  

LAB HEAD: Xuesong Sun

PROVIDER: PXD011244 | Pride | 2022-02-28

REPOSITORIES: Pride

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Improved SILAC method for double labeling of bacterial proteome.

Han Junlong J   Yi Shuhong S   Zhao Xinlu X   Zheng Yundan Y   Yang Donghong D   Du Gaofei G   Yang Xiao-Yan XY   He Qing-Yu QY   Sun Xuesong X  

Journal of proteomics 20181213


Stable isotope labeling with amino acids in cell culture (SILAC) is a robust proteomics method with advantages such as reproducibility and easy handling. This method is popular for the analysis of mammalian cells. However, amino acid conversion in bacteria decreases the labeling efficiency and quantification accuracy, limiting the application of SILAC in bacterial proteomics to auxotrophic bacteria or to single labeling with lysine. In this study, we found that adding high concentrations of isot  ...[more]

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