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Sortagging: a versatile method for protein labeling

Nature Chemical Biology volume 3, pages 707–708 (2007)Cite this article

Abstract

Genetically encoded reporter constructs that yield fluorescently labeled fusion proteins are a powerful tool for observing cell biological phenomena, but they have limitations. Sortagging (sortase-mediated transpeptidation) is a versatile chemoenzymatic system for site-specific labeling of proteins with small (<2 kDa) probes. Sortagging combines the precision of a genetically encoded tag with the specificity of an enzymatic reaction and the ease and chemical versatility of peptide synthesis. Here we apply this technique to proteins in vitro and on the surface of living cells.

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Figure 1: Probes compatible with sortagging and quantitative labeling of H-2Kb complexes in vitro.
Figure 2: Labeling of proteins on cell surfaces.

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Acknowledgements

We thank O. Schneewind for the sortase A–encoding plasmid, C.H. Shen and J. Chen for the neuraminidase template, and the Ploegh lab and H. Hang for helpful discussions. This work was supported by grants from the US National Institutes of Health.

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Authors and Affiliations

  1. Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, 02142, Massachusetts, USA

    Maximilian W Popp, John M Antos, Gijsbert M Grotenbreg, Eric Spooner & Hidde L Ploegh

  2. Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02142, Massachusetts, USA

    Maximilian W Popp & Hidde L Ploegh

Authors
  1. Maximilian W Popp
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  2. John M Antos
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  3. Gijsbert M Grotenbreg
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  4. Eric Spooner
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  5. Hidde L Ploegh
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Contributions

M.W.P., J.M.A. and G.M.G. performed experiments and prepared the manuscript; E.S. performed mass spectrometry analysis; H.L.P. prepared the manuscript and conceived of the project.

Corresponding author

Correspondence to Hidde L Ploegh.

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Supplementary Figures 1–6, Supplementary Table 1 and Supplementary Methods (PDF 1193 kb)

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Popp, M., Antos, J., Grotenbreg, G. et al. Sortagging: a versatile method for protein labeling. Nat Chem Biol 3, 707–708 (2007). https://doi.org/10.1038/nchembio.2007.31

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