Site-specific N-terminal labeling of proteins using sortase-mediated reactions
Author(s)Theile, Christopher S.; Witte, Martin D.; Blom, Annet E. M.; Kundrat, Lenka; Guimaraes, Carla P.; Ploegh, Hidde; ... Show more Show less
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This protocol describes the use of sortase-mediated reactions to label the N terminus of any given protein of interest. The sortase recognition sequence, LPXTG (for Staphylococcus aureus sortase A) or LPXTA (for Streptococcus pyogenes sortase A), can be appended to a variety of probes such as fluorophores, biotin or even to other proteins. The protein to be labeled acts as a nucleophile by attacking the intermediate formed between the probe containing the LPXTG/A motif and the sortase enzyme. If sortase, the protein of interest and a suitably functionalized label are available, the reactions usually require less than 3 h.
DepartmentMassachusetts Institute of Technology. Department of Biology; Whitehead Institute for Biomedical Research
Nature Publishing Group
Theile, Christopher S, Martin D Witte, Annet E M Blom, Lenka Kundrat, Hidde L Ploegh, and Carla P Guimaraes. “Site-Specific N-Terminal Labeling of Proteins Using Sortase-Mediated Reactions.” Nat Protoc 8, no. 9 (August 29, 2013): 1800–1807.