Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations
Purchase on Springer.com
$39.95 / €34.95 / £29.95*
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.
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.
- Bruix, M., Jimenez-Barbero, J. and Cronet, P. (1995) Carbohydr. Res., 273, 157–170.
- Erbel, P.J., Barr, K., Gao, N., Gerwig, G.J., Rick, P.D. and Gardner, K.H. (2003) J. Bacteriol., 185, 1995–2004.
- Kahn, R.A., Randazzo, P., Serafini, T., Weiss, O., Rulka, C., Clark, J., Amherdt, M., Roller, P., Orci, L. and Rothman, J.E. (1992) J. Biol. Chem., 267, 13039–46.
- Lugowski, C., Romanowska, E., Kenne, L. and Lindberg, B. (1983) Carbohydr. Res., 118, 173–181.
- MacIntosh, S.E. (2001) M.Sc. Thesis, University of British Columbia, Vancouver.
- Mackereth, C.D., Scharpf, M., Gentile, L.N. and McIntosh, L.P. (2002) J. Biomol. NMR, 24, 71–2.
- Neidhardt, F.C., Bloch, P.L. and Smith, D.F. (1974) J. Bacteriol., 119, 736–747.
- Parsons, L., Bonander, N., Eisenstein, E., Gilson, M., Kairys, V. and Orban, J. (2003) Biochemistry, 42, 80–89.
- Rick, P.D. and Silver, R.P. (1996) In Escherichia coli and Salmonella: Cellular and Molecular Biology, Vol. 1, Neidhardt, F.C. (Ed.), American Society for Microbiology, Washington, D.C., pp. 104–122.
- Staaf, M., Hoog, C., Stevensson, B., Maliniak, A. and Widmalm, G. (2001) Biochemistry, 40, 3623–8.
- Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations
Journal of Biomolecular NMR
Volume 29, Issue 2 , pp 199-204
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- enterobacterial common antigen
- heterologous protein expression
- N-acetylated carbohydrate
- protein purification
- Industry Sectors
- Author Affiliations
- 1. Departments of Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX, 75390-9038, U.S.A.
- 2. Complex Carbohydrate Research Center, University of Georgia, Athens, GA, 30602-4712, U.S.A.
- 3. Department of Biochemistry and Molecular Biology, Department of Chemistry, and the Protein Engineering Network of Centres of Excellence, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada
- 4. Department of Biochemistry, Emory University School of Medicine, Atlanta, GA, 30322, U.S.A.