Martin D Witte, Tongfei Wu, Carla P Guimaraes, Christopher S Theile, Annet E M Blom, Jessica R Ingram, Zeyang Li, Lenka Kundrat, Shalom D Goldberg, Hidde L Ploegh
Transpeptidation catalyzed by sortase A allows the preparation of proteins that are site-specifically and homogeneously modified with a wide variety of functional groups, such as fluorophores, PEG moieties, lipids, glycans, bio-orthogonal reactive groups and affinity handles. This protocol describes immobilization of sortase A on a solid support (Sepharose beads). Immobilization of sortase A simplifies downstream purification of a protein of interest after labeling of its N or C terminus. Smaller batch and larger-scale continuous-flow reactions require only a limited amount of enzyme...
March 2015: Nature Protocols