Stable Isotope Labeling of a Group a Streptococcus Virulence Factor Using a Chemically Defined Growth Medium

Authors

Pamela D. Vise, University of Idaho
Kishore Kodali, University of Idaho
Nancy Hoe, National Institute of Health
Andrzej Paszczynski, University of Idaho
James M. Musser, Baylor College of Medicine
Gary W. Daughdrill, University of Idaho

Document Type

Article

Publication Date

12-2003

Digital Object Identifier (DOI)

https://doi.org/10.1016/S1046-5928(03)00235-3

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 ¹⁵ N, at the amide nitrogen. The ¹⁵ N-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.

Was this content written or created while at USF?

No

Citation / Publisher Attribution

Protein Expression and Purification, v.32, issue 2, p. 232-238

Scholar Commons Citation

Vise, Pamela D.; Kodali, Kishore; Hoe, Nancy; Paszczynski, Andrzej; Musser, James M.; and Daughdrill, Gary W., "Stable Isotope Labeling of a Group a Streptococcus Virulence Factor Using a Chemically Defined Growth Medium" (2003). Molecular Biosciences Faculty Publications. 143.
https://digitalcommons.usf.edu/bcm_facpub/143