TL;DR
The transpeptidation of two nanobodies (VHHs, variable region of the heavy chain of heavy chain of heavy-chain only antibodies) with various GGG-nucleophiles is described.
The potential of BpyAla-mediated nucleic acid cleavage is discussed, with emphasis on the development of next-generation BpyAla analogues, the exploration of alternative metal cofactors, cooperative and multi-residue design strategies, and the expansion of compatible protein scaffolds and nucleic acid substrates.
Bacterial sortases are widely used in sortase-mediated ligation (SML) experiments for various protein engineering applications. The power of these enzymes to bind and cleave a specific recognition motif, followed by ligation to another substrate using a ping-pong reaction mechanism has numerous applications in vaccine...
Sortase A (SrtA) enzymes covalently anchor surface proteins to Gram-positive bacterial cell walls, promoting colonization and virulence. In Streptococcus pneumoniae, previous studies identified both a domain-swapped dimer and an active refolded monomer, but the active enzyme has not been characterized at the structural...
The development of site-specific artificial nucleases remains a significant challenge in chemical biology. Here, we report the design and biophysical characterization of a light-responsive protein-small molecule hybrid based on the EL222 transcription factor. Through protein bioconjugation, we converted the native LOV-...
NMR data, in combination with enzymatic assays using active variants confirmed differences in the active, apo states of these enzymes, and provided additional atomic detail regarding the importance of the P94 residue in saSrtA substrate recognition.
The inclusion of non-proteinogenic amino acids (npAAs) into proteins vastly expands their chemical repertoire and hence possible functional diversity. However, it remains challenging to obtain non-proteinogenic sequences that carry out a specific task. Here we develop a platform to discover functional non-standard prot...
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