Enhancing Robustness of Sortase A by Loop Engineering and Backbone Cyclization.

Zhi Zou,Ulrich Schwaneberg,Felix Jakob,Maximilian Nöth,Diana M. Mate

doi:10.1002/chem.202004039

Abstract

Invited for the cover of this issue is Ulrich Schwaneberg and co-workers at RWTH Aachen University and DWI Leibniz-Institut für Interaktive Materialien. The image depicts a loop engineered, and backbone cyclized Staphylococcus aureus sortase A which shows enhanced robustness in site-specific protein and peptide modifications. Read the full text of the article at 10.1002/chem.202002740.

Highlights

Functionalization of proteins is often performed by targeting reactive endogenous amino acid side chains (e.g. -NH2 in lysine or -SH in cysteine residues).[1]
The resulting protein-sortase thioester is attacked by the amino group of an N-terminal glycine residue of protein 2 (P2), resulting in a protein conjugate of protein 1 and protein 2 (Scheme 1).[14]
Given to the comparably high transpeptidase activity among the sortases,[21] Staphylococcus aureus sortase A (SaSrtA) is most frequently used in sortagging applications.[22]

Summary

Introduction

Functionalization of proteins is often performed by targeting reactive endogenous amino acid side chains (e.g. -NH2 in lysine or -SH in cysteine residues).[1]. A few studies reported the engineering of SaSrtA WT for increased stability towards thermal and chemical denaturation (e.g. 11.2 8C improvement in melting temperature (Tm) and 4.5-fold resistance in presence of 2.5 m urea).[31,32] engineering of highly active SaSrtA variants with enhanced tolerance against thermal and chemical stress has not been reported yet.

Results

Conclusion