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Protein Semi-Synthesis in Living Cells
Abstract
Incorporation of chemical probes into proteins is a powerful way to elucidate biological processes and to engineer novel function. Here we describe an approach that allows ligation of synthetic molecules to target proteins in an intracellular environment. A cellular protein is genetically tagged with one-half of a split intein. The complementary half is linked in vitro to the synthetic probe, and this fusion is delivered into cells using a transduction peptide. Association of the intein halves in the cytosol triggers protein trans-splicing, resulting in the ligation of the probe to the target protein through a peptide bond. This process is specific and applicable to cytosolic and integral membrane proteins. The technology should allow cellular proteins to be elaborated with a variety of abiotic probes.
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Cited by
Bioorthogonal Chemistry: Fishing for Selectivity in a Sea of Functionality
Sletten, E. M., & Bertozzi, C. R. (2009) · Angewandte Chemie International Edition · doi:10.1002/anie.200900942 · Cited by 3,165
Prescher, J. A., & Bertozzi, C. R. (2005) · Nature Chemical Biology · doi:10.1038/nchembio0605-13 · Cited by 1,456
Bioorthogonale Chemie – oder: in einem Meer aus Funktionalität nach Selektivität fischen
Sletten, E. M., & Bertozzi, C. R. (2009) · Angewandte Chemie · doi:10.1002/ange.200900942 · Cited by 564
High-resolution multi-dimensional NMR spectroscopy of proteins in human cells
Inomata, K., Ohno, A., Tochio, H., Isogai, S., Tenno, T., Nakase, I., Takeuchi, T., Futaki, S., Ito, Y., Hiroaki, H., et al. (2009) · Nature · doi:10.1038/nature07839 · Cited by 458
Inteins: nature's gift to protein chemists
Shah, N. H., & Muir, T. W. (2014) · Chem. Sci. · doi:10.1039/c3sc52951g · Cited by 404
Self-citationProtein ligation: an enabling technology for the biophysical analysis of proteins
Muralidharan, V., & Muir, T. W. (2006) · Nature Methods · doi:10.1038/nmeth886 · Cited by 364
Self-citationChemical Synthesis of Proteins
Nilsson, B. L., Soellner, M. B., & Raines, R. T. (2005) · Annual Review of Biophysics and Biomolecular Structure · doi:10.1146/annurev.biophys.34.040204.144700 · Cited by 329
In vivo protein labeling with trimethoprim conjugates: a flexible chemical tag
Miller, L. W., Cai, Y., Sheetz, M. P., & Cornish, V. W. (2005) · Nature Methods · doi:10.1038/nmeth749 · Cited by 323
Reversible site-selective labeling of membrane proteins in live cells
Guignet, E. G., Hovius, R., & Vogel, H. (2004) · Nature Biotechnology · doi:10.1038/nbt954 · Cited by 291
Histones: At the Crossroads of Peptide and Protein Chemistry
Müller, M. M., & Muir, T. W. (2015) · Chemical Reviews · doi:10.1021/cr5003529 · Cited by 247
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Metadata
- Authors
- Giriat, I., & Muir, T. W.
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2003-05-22
- Volume
- 125
- Issue
- 24
- Pages
- 7180-7181
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W2014718744
- Cited by
- 167
- References
- 0
- FWCI
- 5.47
- Type
- journal article
- Language
- en
- Open access
- closed
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