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Conjugation of Glycopeptide Thioesters to Expressed Protein Fragments

Semisynthesis of Glycosylated Interleukin-2
  • Thomas J. Tolbert
  • Chi-Huey Wong
Part of the Methods in Molecular Biology™ book series (MIMB, volume 283)

Abstract

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.

Key Words

Glycoprotein chemoselective native chemical ligation expressed protein ligation semisynthesis glycopeptide thioester conjugation interleukin-2 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Thomas J. Tolbert
    • 1
  • Chi-Huey Wong
    • 2
  1. 1.Department of ChemistryIndiana UniversityBloomington
  2. 2.Department of ChemistryThe Scripps Research InstituteLa Jolla

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