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Thioester-Assisted Sortase-A-Mediated Ligation.

Chong Zuo1, Ruichao Ding1, Xiangwei Wu1

  • 1Tsinghua-Peking Center for Life Sciences, Ministry of Education Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, State Key Laboratory of Chemical Oncogenomics (Shenzhen), Department of Chemistry, Tsinghua University, Beijing, 100084, China.

Angewandte Chemie (International Ed. in English)
|May 6, 2022
PubMed
Summary

Sortase A (SrtA) ligation is now more efficient and irreversible using thioester-assisted methods. This protein labeling technique broadens substrate compatibility and simplifies the preparation of modified proteins.

Keywords:
Chemical Protein SynthesisProtein LabellingProtein Semi-SynthesisSortase-A-Mediated LigationThioesters

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Molecular Biology

Background:

  • Sortase A (SrtA)-mediated ligation is a widely used technique for protein labeling and semi-synthesis.
  • Current limitations include reaction reversibility and strict dependence on the LPxTG recognition motif.

Purpose of the Study:

  • To develop a more efficient and irreversible Sortase A ligation method.
  • To expand the utility of Sortase A ligation beyond the traditional LPxTG motif.

Main Methods:

  • Developed a thioester-assisted Sortase A ligation strategy.
  • Utilized proteins/peptides with C-terminal thioesters and N-terminal Glycine.
  • Tested ligation with various LPxT-derived sequences and post-translationally modified proteins.

Main Results:

  • Achieved efficient and irreversible Sortase A ligation using the thioester-assisted method.
  • Demonstrated broad tolerance for diverse LPxT-derived sequences.
  • Successfully prepared proteins with N- or C-terminal labels, including phosphorylated and methylated histones.

Conclusions:

  • Chemical modification of substrates enhances the synthetic capabilities of enzymatic ligation methods.
  • The thioester-assisted SrtA ligation offers a versatile approach for protein labeling and semi-synthesis.
  • This method reduces dependence on the canonical LPxTG motif for Sortase A-mediated reactions.