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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: May 15, 2026

Synthesis of an Intein-mediated Artificial Protein Hydrogel
15:06

Synthesis of an Intein-mediated Artificial Protein Hydrogel

Published on: January 27, 2014

Streamlined expressed protein ligation using split inteins.

Miquel Vila-Perelló1, Zhihua Liu, Neel H Shah

  • 1Department of Chemistry, Princeton University, Frick Laboratory, Princeton, New Jersey 08544, United States.

Journal of the American Chemical Society
|December 26, 2012
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method using engineered split DnaE inteins for efficient preparation of protein alpha-thioesters. This advancement simplifies the generation of modified proteins for research and therapeutic applications.

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Last Updated: May 15, 2026

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Chemically modified proteins are crucial for biological studies and therapeutics.
  • Expressed protein ligation (EPL) is a common method for protein modification, but requires challenging protein alpha-thioester generation.
  • Existing methods for recombinant protein alpha-thioester synthesis are often complex and inefficient.

Purpose of the Study:

  • To develop a streamlined and efficient method for preparing recombinant protein alpha-thioesters.
  • To utilize engineered split DnaE inteins for improved protein modification strategies.
  • To demonstrate the application of this new method in protein semisynthesis.

Main Methods:

  • Engineered naturally split DnaE inteins were used for protein alpha-thioester preparation.
  • A purification strategy leveraging the intein fragments' affinity was developed.
  • The method was applied to the semisynthesis of modified proteins, including acetylated histone and monoclonal antibodies.

Main Results:

  • A new, streamlined procedure for generating and purifying protein alpha-thioesters from cell lysates was established.
  • Engineered split DnaE inteins demonstrated faster kinetics compared to existing inteins.
  • Successful semisynthesis of complex proteins, such as modified antibodies and histones, was achieved.

Conclusions:

  • The developed method offers an efficient alternative for producing protein alpha-thioesters.
  • This technique simplifies protein modification and expands its utility in biological research and drug development.
  • Engineered split DnaE inteins represent a powerful tool for advancing protein engineering and therapeutic applications.