Stable Isotope Labeling by Amino Acids Applied to Bacterial Cell Culture
Stable isotope labeling by amino acids in cell culture (SILAC) is a widely used approach in quantitative proteomics; however, due to limitations such as required auxotrophy for the amino acids employed for labeling, it was thus far rarely employed in bacteria. Although limitations of SILAC in microbiological applications are significant and restrict its use exclusively to cells cultured in minimal media, we and others have successfully used it to fully label proteomes of model bacteria and measure their relative expression dynamics under different experimental conditions. Here we provide a brief overview of applications of SILAC in bacteria and describe a detailed protocol for SILAC labeling of Escherichia coli and Bacillus subtilis cells in culture, which in many cases can be applied to other members of both gram-positive and gram-negative bacterial species.
Key wordsSILAC Quantitative proteomics Mass spectrometry Bacteria Prokaryotes
The authors wish to thank Alejandro Carpy and the other PCT members for useful comments on the manuscript. Our work is financed by the Juniorprofessoren-Programm of the Landesstiftung BW, the SFB766 of the Deutsche Forschungsgemeinschaft, and PRIME-XS consortium.
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