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

Pamela D. Vise, Kishore Kodali, Nancy Hoe, Andrzej Paszczynski, James M. Musser, Gary W. Daughdrill

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)232-238
Number of pages7
JournalProtein Expression and Purification
Volume32
Issue number2
DOIs
StatePublished - Dec 2003

ASJC Scopus subject areas

  • Biotechnology

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