Conjugation of glycopeptide thioesters to expressed protein fragments: semisynthesis of glycosylated interleukin-2.
This method describes the conjugation of a synthetic glycopeptide to the N-terminus of a recombinant human interleukin-2 (IL-2) protein fragment. The IL-2 protein fragment is produced as an affinity-tagged fusion protein in Escherichia coli and then cleaved with the highly selective TEV protease to remove the affinity tag and uncover an N-terminal cysteine. The N-terminal cysteine is then used in native chemical ligation reaction to join the IL-2 protein fragment to a glycosylated tripeptide thioester that had been previously synthesized to produce a glycosylated form of IL-2.
Tolbert
Thomas J
TJ
Department of Chemistry, Indiana University, Bloomington, USA.
Wong
Chi-Huey
CH
D003545:Cysteine / Q000138:chemical synthesis / Q000235:genetics; D010450:Endopeptidases; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis / Q000138:chemical synthesis / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics
D006020:Glycopeptides; D007376:Interleukin-2; D010455:Peptides; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; C099488:TEV protease; D003545:Cysteine