Making and Breaking Peptide Bonds: Protein Engineering Using Sortase

• Published in: Angewandte Chemie (International ed.) Vol. 50; no. 22; pp. 5024 - 5032
• Main Authors: Popp, Maximilian Wei-Lin, Ploegh, Hidde L.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 23.05.2011; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Edition: International ed. in English
• Subjects: Amino Acid Motifs; Aminoacyltransferases - chemistry; Aminoacyltransferases - genetics; Aminoacyltransferases - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - genetics; Cysteine Endopeptidases - metabolism; Enzymes; Protein Engineering; Protein folding; protein modifications; Protein Structure, Tertiary; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; site specificity; sortase; Staphylococcus aureus - enzymology; transpeptidation
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201008267

Sortases are a class of bacterial enzymes that possess transpeptidase activity. It is their ability to site‐specifically break a peptide bond and then reform a new bond with an incoming nucleophile that makes sortase an attractive tool for protein engineering. This technique has been adopted for a range of applications, from chemistry‐based to cell biology and technology. In this Minireview we provide a brief overview of the biology of sortase enzymes and current applications in protein engineering. We identify areas that lend themselves to further innovation and that suggest new applications. It takes all sortases: Enzymatic formation of a peptide bond using the sortase A transpeptidase (SrtA) provides a convenient and mild means for engineering proteins to contain nongenetically templated modifications. Myriad applications are possible, from producing homogeneous post‐translational modification mimics, assembling protein domains, to anchoring proteins to solid surfaces.