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Related Experiment Video

Updated: Feb 19, 2026

Detection of Detergent-sensitive Interactions Between Membrane Proteins

Detection of Detergent-sensitive Interactions Between Membrane Proteins

Published on: March 7, 2018

Expanding the Scope of Sortase-Mediated Ligations by Using Sortase Homologues.

Keyvan D Nikghalb¹, Nicholas M Horvath¹, Jesse L Prelesnik¹

¹Department of Chemistry, Western Washington University, 516 High Street, Bellingham, WA, 98225, USA.

Chembiochem : a European Journal of Chemical Biology

|November 11, 2017

Summary

Researchers expanded the utility of sortase enzymes for creating novel protein derivatives. They identified new sortase A variants with broader substrate recognition, enabling new protein modification strategies.

Keywords:

enzymes peptides protein modifications sortase transacylation

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Area of Science:

Biochemistry
Molecular Biology
Protein Engineering

Background:

Sortase-catalyzed transacylation is crucial for synthesizing non-natural protein derivatives.
The commonly used Sortase A from *Staphylococcus aureus* has a limited substrate scope, hindering broader applications.

Purpose of the Study:

To identify sortase A homologues with expanded substrate recognition capabilities.
To develop novel protein modification strategies using engineered sortase enzymes.

Main Methods:

In vitro characterization of substrate preferences for eight sortase A homologues.
Site-specific modification of the antimicrobial peptide DCD-1L using *Streptococcus pneumoniae* sortase A.
Analysis of alternative transacylation sites and oligomer formation.

Main Results:

Identified sortase A enzymes with broader substrate scope than *S. aureus* Sortase A.
Demonstrated site-specific N-terminal modification of DCD-1L using *S. pneumoniae* Sortase A.
Observed unexpected alternative transacylation sites and oligomer formation with specific substrates.

Conclusions:

This study expands the toolkit of sortase enzymes for protein engineering.
The identified sortase variants and observed novel reactivities pave the way for advanced protein modification techniques.
Further development of sortase enzymes holds significant promise for creating diverse protein derivatives.