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

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)199-204
Number of pages6
JournalJournal of Biomolecular NMR
Volume29
Issue number2
DOIs
StatePublished - Jun 2004

Fingerprint

Nuclear magnetic resonance
Impurities
Carbohydrates
Proteins
Escherichia coli
Polysaccharides
Water
enterobacterial common antigen

Keywords

  • Enterobacterial common antigen
  • Heterologous protein expression
  • N-acetylated carbohydrate
  • Protein purification

ASJC Scopus subject areas

  • Spectroscopy
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Cyclic enterobacterial common antigen : Potential contaminant of bacterially expressed protein preparations. / Erbel, Paul J A; Seidel, Ronald; Macintosh, Scott E.; Gentile, Lisa N.; Amor, Juan C.; Kahn, Richard A.; Prestegard, James H.; McIntosh, Lawrence P.; Gardner, Kevin H.

In: Journal of Biomolecular NMR, Vol. 29, No. 2, 06.2004, p. 199-204.

Research output: Contribution to journalArticle

Erbel, PJA, Seidel, R, Macintosh, SE, Gentile, LN, Amor, JC, Kahn, RA, Prestegard, JH, McIntosh, LP & Gardner, KH 2004, 'Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations', Journal of Biomolecular NMR, vol. 29, no. 2, pp. 199-204. https://doi.org/10.1023/B:JNMR.0000019252.65073.24
Erbel, Paul J A ; Seidel, Ronald ; Macintosh, Scott E. ; Gentile, Lisa N. ; Amor, Juan C. ; Kahn, Richard A. ; Prestegard, James H. ; McIntosh, Lawrence P. ; Gardner, Kevin H. / Cyclic enterobacterial common antigen : Potential contaminant of bacterially expressed protein preparations. In: Journal of Biomolecular NMR. 2004 ; Vol. 29, No. 2. pp. 199-204.
@article{24c9166849344ec2ac854275724319a2,
title = "Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations",
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.",
keywords = "Enterobacterial common antigen, Heterologous protein expression, N-acetylated carbohydrate, Protein purification",
author = "Erbel, {Paul J A} and Ronald Seidel and Macintosh, {Scott E.} and Gentile, {Lisa N.} and Amor, {Juan C.} and Kahn, {Richard A.} and Prestegard, {James H.} and McIntosh, {Lawrence P.} and Gardner, {Kevin H.}",
year = "2004",
month = "6",
doi = "10.1023/B:JNMR.0000019252.65073.24",
language = "English (US)",
volume = "29",
pages = "199--204",
journal = "Journal of Biomolecular NMR",
issn = "0925-2738",
publisher = "Springer Netherlands",
number = "2",

}

TY - JOUR

T1 - Cyclic enterobacterial common antigen

T2 - Potential contaminant of bacterially expressed protein preparations

AU - Erbel, Paul J A

AU - Seidel, Ronald

AU - Macintosh, Scott E.

AU - Gentile, Lisa N.

AU - Amor, Juan C.

AU - Kahn, Richard A.

AU - Prestegard, James H.

AU - McIntosh, Lawrence P.

AU - Gardner, Kevin H.

PY - 2004/6

Y1 - 2004/6

N2 - 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.

AB - 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.

KW - Enterobacterial common antigen

KW - Heterologous protein expression

KW - N-acetylated carbohydrate

KW - Protein purification

UR - http://www.scopus.com/inward/record.url?scp=1842607683&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842607683&partnerID=8YFLogxK

U2 - 10.1023/B:JNMR.0000019252.65073.24

DO - 10.1023/B:JNMR.0000019252.65073.24

M3 - Article

VL - 29

SP - 199

EP - 204

JO - Journal of Biomolecular NMR

JF - Journal of Biomolecular NMR

SN - 0925-2738

IS - 2

ER -