Generation of Ca2+-independent sortase A mutants with enhanced activity for protein and cell surface labeling
2017; 12 (12): e0189068
Sortase A, a calcium-dependent transpeptidase derived from Staphylococcus aureus, is used in a broad range of applications, such as the conjugation of fluorescent dyes and other moieties to proteins or to the surface of eukaryotic cells. In vivo and cell-based applications of sortase have been somewhat limited by the large range of calcium concentrations, as well as by the often transient nature of protein-protein interactions in living systems. In order to use sortase A for cell labeling applications, we generated a new sortase A variant by combining multiple mutations to yield an enzyme that was both calcium-independent and highly active. This variant has enhanced activity for both N- and C-terminal labeling, as well as for cell surface modification under physiological conditions.
View details for DOI 10.1371/journal.pone.0189068
View details for Web of Science ID 000417033200027
View details for PubMedID 29200433
View details for PubMedCentralID PMC5714338