Stable isotope labeling of a Group A Streptococcus virulence factor using a chemically defined growth medium

A secreted, hypervariable virulence factor called the streptococcal inhibitor of complement (Sic) has been linked to the reemergence of epidemics due to the human pathogenic bacterium Group A Streptococcus. This paper describes a method for expressing and purifying Sic from an attenuated GAS strain...

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Veröffentlicht in:Protein expression and purification 2003-12, Vol.32 (2), p.232-238
Hauptverfasser: Vise, Pamela D, Kodali, Kishore, Hoe, Nancy, Paszczynski, Andrzej, Musser, James M, Daughdrill, Gary W
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Sprache:eng
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Zusammenfassung:A secreted, hypervariable virulence factor called the streptococcal inhibitor of complement (Sic) has been linked to the reemergence of epidemics due to the human pathogenic bacterium Group A Streptococcus. This paper describes a method for expressing and purifying Sic from an attenuated GAS strain using a chemically defined growth medium. This method was used to label specific amino acid residue types in Sic with forms containing the magnetically active isotope 15N, at the amide nitrogen. The 15N-labeling of Sic permits a detailed investigation of the structure and dynamics of the protein using nuclear magnetic resonance spectroscopy. The level of stable isotope incorporation was established using mass spectrometry and nuclear magnetic resonance spectroscopy.
ISSN:1046-5928
1096-0279
DOI:10.1016/S1046-5928(03)00235-3