Related Experiment Video
Updated: Jul 7, 2026

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli
Published on: December 9, 2017
Genetically encoding N(epsilon)-acetyllysine in recombinant proteins.
Heinz Neumann1, Sew Y Peak-Chew, Jason W Chin
1Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH England, UK.
Scientists developed a new method to insert N(epsilon)-acetyllysine into proteins at specific sites. This breakthrough in protein synthesis will help researchers understand the role of this important post-translational modification.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- N(epsilon)-acetylation of lysine is a crucial reversible post-translational modification in eukaryotes.
- This modification's regulatory impact is comparable to that of phosphorylation.
- Currently, no general methods exist for the site-specific synthesis of proteins containing N(epsilon)-acetyllysine.
Purpose of the Study:
- To develop a general method for the site-specific incorporation of N(epsilon)-acetyllysine into recombinant proteins.
- To enable the study of N(epsilon)-acetyllysine's function in biological systems.
Main Methods:
- Evolution of an orthogonal N(epsilon)-acetyllysyl-tRNA synthetase/tRNA(CUA) pair.
- Site-specific incorporation of N(epsilon)-acetyllysine in recombinant proteins produced in Escherichia coli.
Main Results:
- Successful demonstration of site-specific N(epsilon)-acetyllysine incorporation in recombinant proteins.
- Establishment of a novel protein synthesis strategy using an engineered tRNA synthetase/tRNA pair.
Conclusions:
- The developed method provides a powerful tool for synthesizing proteins with site-specific N(epsilon)-acetylation.
- This technique is expected to significantly advance the understanding of N(epsilon)-acetyllysine's cellular roles and regulatory functions.
More Related Videos
07:26Site Specific Lysine Acetylation of Histones for Nucleosome Reconstitution using Genetic Code Expansion in Escherichia coli
Published on: December 26, 2020
11:47Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
Published on: August 1, 2016
Related Concept Videos
Phase II Reactions: Acetylation Reactions
The substrates for acetylation are typically drugs or their metabolites with an amino, sulfonamide, or hydrazine functional group. Acetylation can occur at several points in the drug molecule, including primary, secondary, and...
Covalently Linked Protein Regulators
These groups modify specific amino acids in a protein.
Amines to Amides: Acylation of Amines
Next, the second equivalent of amine serves as a Brønsted base and deprotonates the quaternary amide...
Alkylation of β-Ketoester Enolates: Acetoacetic Ester Synthesis
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...