Labeling, detection and identification of newly synthesized proteomes with bioorthogonal non-canonical amino-acid tagging

@article{Dieterich2007LabelingDA,  title={Labeling, detection and identification of newly synthesized proteomes with bioorthogonal non-canonical amino-acid tagging},  author={Daniela C. Dieterich and Jennifer Lee and A. James Link and Johannes Graumann and David A. Tirrell and Erin Margaret Schuman},  journal={Nature Protocols},  year={2007},  volume={2},  pages={532-540},  url={https://api.semanticscholar.org/CorpusID:2833184}}
This protocol describes the step-by-step labeling, purification and detection of newly synthesized proteins in mammalian cells using the non-canonical amino acid azidohomoalanine (AHA) to identify specific subproteomes.

284 Citations

Non-canonical amino acid labeling in proteomics and biotechnology

This review focuses on techniques in which proteins are residue- and site-specifically labeled with ncAAs containing bioorthogonal handles, and provides decision trees to help guide the design of future experiments.

Quantitative analysis of newly synthesized proteins

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This work demonstrates for the first time the use of a bioorthogonal amino acid for proteome-wide detection of changes in the amounts of proteins synthesized during a brief period upon variations in cellular growth conditions.

Enhancing Comprehensive Analysis of Newly Synthesized Proteins Based on Cleavable Bioorthogonal Tagging.

A novel method for enriching newly synthesized peptides by click chemistry followed by release of enriched peptides via tryptic digestion based on cleavable bioorthogonal tagging (CBOT) that can quantify NSPs when coupling a pair of isotope-labeled azidohomoalanine with decent reproducibility is designed.

Bio-orthogonal labeling as a tool to visualize and identify newly synthesized proteins in Caenorhabditis elegans

This protocol circumvents this limitation by identifying de novo–synthesized proteins via the incorporation of the chemically modifiable azidohomoalanine instead of the natural amino acid methionine in the nascent protein, followed by facilitating the visualization of the resulting labeled proteins in situ.

Click Chemistry (CuAAC) and Detection of Tagged de novo Synthesized Proteins in Drosophila.

Detailed protocols for FUNCAT and BONCAT click chemistry and the detection of tagged de novo synthesized proteins in Drosophila melanogaster are provided.
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