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Updated: May 21, 2026

Constructing Thioether/Vinyl Sulfide-tethered Helical Peptides Via Photo-induced Thiol-ene/yne Hydrothiolation
11:09

Constructing Thioether/Vinyl Sulfide-tethered Helical Peptides Via Photo-induced Thiol-ene/yne Hydrothiolation

Published on: August 1, 2018

Protein thioester synthesis enabled by sortase.

Jingjing J Ling1, Rocco L Policarpo, Amy E Rabideau

  • 1Department of Chemistry, Massachusetts Institute of Technology, 16-573a, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.

Journal of the American Chemical Society
|June 13, 2012
PubMed
Summary
This summary is machine-generated.

Researchers developed a new, simple method using sortase A to create protein thioesters, crucial for protein semisynthesis. This technique facilitates advanced protein engineering and ligation strategies.

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Area of Science:

  • Biochemistry
  • Protein Engineering
  • Synthetic Biology

Background:

  • Proteins with C-terminal thioesters are key intermediates in protein semisynthesis.
  • Current methods for synthesizing protein thioesters primarily rely on engineered inteins.

Purpose of the Study:

  • To introduce a straightforward and routine strategy for preparing recombinant proteins with C-terminal (α)thioesters.
  • To demonstrate the utility of this method in synthesizing complex protein constructs.

Main Methods:

  • Utilized sortase A for the preparation of recombinant proteins containing a C-terminal (α)thioester.
  • Applied the method to synthesize two anthrax toxin cargo proteins, one with a (α)thioester and another with a D-polypeptide segment.
  • Assessed the translocation of these variants through the protective antigen pore.

Main Results:

  • Successfully prepared recombinant proteins with C-terminal (α)thioesters using sortase A.
  • Demonstrated the synthesis of a protein containing a D-polypeptide segment between domains.
  • Confirmed that both engineered protein variants could translocate through the protective antigen pore.

Conclusions:

  • The sortase A-based strategy offers a simple and efficient alternative for synthesizing protein thioesters.
  • This method enables the integration of sortase-mediated ligation with native chemical ligation for advanced protein construction.
  • The developed technique expands possibilities in protein semisynthesis and engineering complex biomolecules.