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Journal of Biomolecular NMR

, Volume 29, Issue 2, pp 199–204 | Cite as

Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations

  • Paul J.A. Erbel
  • Ronald Seidel
  • Scott E. Macintosh
  • Lisa N. Gentile
  • Juan C. Amor
  • Richard A. Kahn
  • James H. Prestegard
  • Lawrence P. McIntosh
  • Kevin H. Gardner
Article

Abstract

We have previously reported the identification of the cyclic enterobacterial common antigen (ECACYC) polysaccharide in E. coli strains commonly used for heterologous protein expression (PJA Erbel et al., J. Bacteriol.185 (2003): 1995). Following this initial report, interactions among several NMR groups established that characteristic N-acetyl signals of ECACYC have been observed in 15N-1H HSQC spectra of samples of various bacterially-expressed proteins suggesting that this water-soluble carbohydrate is a common contaminant. We provide NMR spectroscopic tools to recognize ECACYC in protein samples, as well as several methods to remove this contaminant. Early recognition of ECA-based NMR signals will prevent time-consuming analyses of this copurifying carbohydrate.

enterobacterial common antigen heterologous protein expression N-acetylated carbohydrate protein purification 

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Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Paul J.A. Erbel
    • 1
  • Ronald Seidel
    • 2
  • Scott E. Macintosh
    • 3
  • Lisa N. Gentile
    • 3
  • Juan C. Amor
    • 4
  • Richard A. Kahn
    • 4
  • James H. Prestegard
    • 4
  • Lawrence P. McIntosh
    • 3
  • Kevin H. Gardner
    • 1
  1. 1.Departments of Biochemistry and PharmacologyUniversity of Texas Southwestern Medical CenterDallasU.S.A.
  2. 2.Complex Carbohydrate Research CenterUniversity of GeorgiaAthensU.S.A.
  3. 3.Department of Biochemistry and Molecular Biology, Department of Chemistry, and the Protein Engineering Network of Centres of ExcellenceUniversity of British ColumbiaVancouverCanada
  4. 4.Department of BiochemistryEmory University School of MedicineAtlantaU.S.A.

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