Cell-selective metabolic labeling of proteins

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Abstract

Metabolic labeling of proteins with the methionine surrogate azidonorleucine can be targeted exclusively to specified cells through expression of a mutant methionyl-tRNA synthetase (MetRS). In complex cellular mixtures, proteins made in cells that express the mutant synthetase can be tagged with affinity reagents (for detection or enrichment) or fluorescent dyes (for imaging). Proteins made in cells that do not express the mutant synthetase are neither labeled nor detected.

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Figure 1: Cell-selective labeling of proteomes with azidonorleucine.
Figure 2: Cell-selective protein labeling in mixed populations.
Figure 3: Cell-selective labeling in mixtures of bacterial and mammalian cells.

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Acknowledgements

We gratefully acknowledge the US National Institutes of Health (R21 DA020589 and R01 GM62523) and the US Army Research Office Institute for Collaborative Biotechnologies for support of this work. We thank K. Boulware for assistance in generating the three-dimensional movie of infected macrophages. E.M.S. is an investigator of the Howard Hughes Medical Institute.

Author information

Correspondence to David A Tirrell.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Methods (PDF 2114 kb)

Supplementary Movie 1

Three-dimensional rendering of cell-selective labelling of macrophage infection with E. coli. (MOV 2075 kb)

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Ngo, J., Champion, J., Mahdavi, A. et al. Cell-selective metabolic labeling of proteins. Nat Chem Biol 5, 715–717 (2009) doi:10.1038/nchembio.200

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