Direct visualization of newly synthesized target proteins in situ
Abstract
Protein synthesis is a dynamic process that tunes the cellular proteome in response to internal and external demands. Metabolic labeling approaches identify the general proteomic response but cannot visualize specific newly synthesized proteins within cells. Here we describe a technique that couples noncanonical amino acid tagging or puromycylation with the proximity ligation assay to visualize specific newly synthesized proteins and monitor their origin, redistribution and turnover in situ.
Additional Information
© 2015 Macmillan Publishers Limited. Received 28 April 2014; Accepted 29 January 2015; Published online 16 March 2015. We thank N. Fuerst and A. Staab for the preparation of cultured hippocampal neurons. E.M.S. is funded by the Max Planck Society; an Advanced Investigator award from the European Research Council, Deutsche Forschungsgemeinschaft (DFG) Collaborative Research Center (CRC) 902: Molecular Principles of RNA-based Regulation; DFG CRC 1080: Molecular and Cellular Mechanisms of Neural Homeostasis; and the DFG Cluster of Excellence for Macromolecular Complexes, Goethe University, Frankfurt. D.A.T is funded by US National Institutes of Health grant R01 GM062523.Attached Files
Published - nmeth.3319-SF1.jpg
Accepted Version - nihms676325.pdf
Supplemental Material - nmeth.3319-S1.pdf
Supplemental Material - nmeth.3319-SF2.jpg
Supplemental Material - nmeth.3319-SF3.jpg
Supplemental Material - nmeth.3319-SF4.jpg
Supplemental Material - nmeth.3319-SF5.jpg
Supplemental Material - nmeth.3319-SF6.jpg
Supplemental Material - nmeth.3319-SF7.jpg
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Additional details
- PMCID
- PMC4414919
- Eprint ID
- 54465
- DOI
- 10.1038/nmeth.3319
- Resolver ID
- CaltechAUTHORS:20150205-174152503
- Max Planck Society
- European Research Council (ERC)
- CRC 902: Molecular Principles of RNA-based Regulation
- Deutsche Forschungsgemeinschaft (DFG)
- CRC 1080: Molecular and Cellular Mechanisms of Neural Homeostasis
- Deutsche Forschungsgemeinschaft (DFG)
- R01 GM062523
- NIH
- Created
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2015-03-17Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field