Locked by Design: A Conformationally Constrained Transglutaminase Tag Enables Efficient Site-Specific Conjugation

Angew Chem Int Ed Engl. 2015 Nov 2;54(45):13420-4. doi: 10.1002/anie.201504851. Epub 2015 Sep 14.

Abstract

Based on the crystal structure of a natural protein substrate for microbial transglutaminase, an enzyme that catalyzes protein crosslinking, a recognition motif for site-specific conjugation was rationally designed. Conformationally locked by an intramolecular disulfide bond, this structural mimic of a native conjugation site ensured efficient conjugation of a reporter cargo to the therapeutic monoclonal antibody cetuximab without erosion of its binding properties.

Keywords: antibodies; bioconjugation; protein engineering; site-specific ligation; transglutaminase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cell Line, Tumor
  • Cetuximab / chemistry*
  • Cetuximab / metabolism
  • Cricetulus
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Humans
  • Models, Molecular
  • Protein Conformation
  • Transglutaminases / chemistry*
  • Transglutaminases / metabolism

Substances

  • Disulfides
  • Transglutaminases
  • Cetuximab