It is demonstrated that sortase A (SrtA) can catalyze efficient coupling of peptides to GPI analogues with a gtycine residue attached to the phosphoethanotamine moiety at the nonreducing end to form GPI-linked peptides. This represents the first chemoenzymatic synthesis of GPI-peptide conjugates and is a proof-of-concept for the potential application of SrtA to the synthesis of more complex GPI-anchored peptides/glycopeptides and GPI-anchored proteins/glycoproteins.