A Metabolic Labeling Strategy for Tracking Protein Synthesis in Complex Biological Systems
- 1University of Toronto
Abstract
Protein synthesis supports most biological processes. In the brain in particular, protein synthesis plays a critical role in physiological and pathological states. Here, we describe Tellurophene-Alkyne Cycloaddition-mediated Amino acid Tagging (TeACAT), a versatile strategy for fast, facile, and flexible tagging of newly synthesized proteins in mice. TeACAT is based on metabolic incorporation of the non-canonical amino acid TePhe into proteins by the endogenous protein synthesis machinery. Due to their high similarity, TePhe can efficiently replace canonical Phe without dietary or genetic manipulation. The subsequent bio-orthogonal reaction of TePhe with either fluorescent dyes or affinity handles enables both visualization and affinity enrichment of proteins synthesized during TePhe exposure. TeACAT is compatible with immunofluorescence for cell-type specific visualization of protein synthesis with subcellular resolution and can be used in conjunction with routine proteomics to identify and quantify newly synthesized proteins. Robust incorporation into the mouse proteome was observed on the scale of hours to days, allowing the interrogation of various biological processes. In summary, TeACAT enables the visualization and quantification of protein synthesis with minimal perturbation for biological discoveries.
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BibTeX
@misc{bu2026metabolic,
title = {{A Metabolic Labeling Strategy for Tracking Protein Synthesis in Complex Biological Systems}},
author = {Y. J. Bu and S. P. Nyandwi and F. De Lima Alves and R. Tennakoon and T. V. Stamm and D. J. Schneider and A. Eddenden and T. W. Y. Ma and Y.-j. Chun and H. Peng and J. M. Miller and A. R. Wheeler and S. Yuzwa and M. Nitz and H. Cui},
howpublished = {bioRxiv},
year = {2026},
month = aug,
doi = {10.64898/2026.08.30.747940},
url = {https://www.biorxiv.org/content/10.64898/2026.08.30.747940v1},
note = {CC BY-ND},
}
RIS
TY - UNPB
TI - A Metabolic Labeling Strategy for Tracking Protein Synthesis in Complex Biological Systems
AU - Y. J. Bu
AU - S. P. Nyandwi
AU - F. De Lima Alves
AU - R. Tennakoon
AU - T. V. Stamm
AU - D. J. Schneider
AU - A. Eddenden
AU - T. W. Y. Ma
AU - Y.-j. Chun
AU - H. Peng
AU - J. M. Miller
AU - A. R. Wheeler
AU - S. Yuzwa
AU - M. Nitz
AU - H. Cui
PY - 2026
DA - 2026/08/30
PB - bioRxiv
DO - 10.64898/2026.08.30.747940
UR - https://www.biorxiv.org/content/10.64898/2026.08.30.747940v1
AB - Protein synthesis supports most biological processes. In the brain in particular, protein synthesis plays a critical role in physiological and pathological states. Here, we describe Tellurophene-Alkyne Cycloaddition-mediated Amino acid Tagging (TeACAT), a versatile strategy for fast, facile, and flexible tagging of newly synthesized proteins in mice. TeACAT is based on metabolic incorporation of the non-canonical amino acid TePhe into proteins by the endogenous protein synthesis machinery. Due to their high similarity, TePhe can efficiently replace canonical Phe without dietary or genetic manipulation. The subsequent bio-orthogonal reaction of TePhe with either fluorescent dyes or affinity handles enables both visualization and affinity enrichment of proteins synthesized during TePhe exposure. TeACAT is compatible with immunofluorescence for cell-type specific visualization of protein synthesis with subcellular resolution and can be used in conjunction with routine proteomics to identify and quantify newly synthesized proteins. Robust incorporation into the mouse proteome was observed on the scale of hours to days, allowing the interrogation of various biological processes. In summary, TeACAT enables the visualization and quantification of protein synthesis with minimal perturbation for biological discoveries.
ER -