title: Mapping the protein-protein interface between a toxin and its cognate antitoxin from the bacterial pathogen Streptococcus pyogenes.
abstract: Protein--protein interactions are ubiquitous and essential for most biological processes. Although new proteomic technologies have generated large catalogs of interacting proteins, considerably less is known about these interactions at the molecular level, information that would aid in predicting protein interactions, designing therapeutics to alter these interactions, and understanding the effects of disease-producing mutations. Here we describe mapping the interacting surfaces of the bacterial toxin SPN (Streptococcus pyogenes NAD(+) hydrolase) in complex with its antitoxin IFS (immunity factor for SPN) by using hydrogen-deuterium amide exchange and electrospray ionization mass spectrometry. This approach affords data in a relatively short time for small amounts of protein, typically 5-7 pmol per analysis. The results show a good correspondence with a recently determined crystal structure of the IFS--SPN complex but additionally provide strong evidence for a folding transition of the IFS protein that accompanies its binding to SPN. The outcome shows that mass-based chemical footprinting of protein interaction surfaces can provide information about protein dynamics that is not easily obtained by other methods and can potentially be applied to large, multiprotein complexes that are out of range for most solution-based methods of biophysical analysis.
authors.affiliation: Analytical Research and Development, Pfizer Inc., Chesterfield, Missouri 63017, United States.
authors.forename: Justin B
authors.initials: JB
authors.lastname: Sperry
authors.forename: Craig L
authors.initials: CL
authors.lastname: Smith
authors.forename: Michael G
authors.initials: MG
authors.lastname: Caparon
authors.forename: Tom
authors.initials: T
authors.lastname: Ellenberger
authors.forename: Michael L
authors.initials: ML
authors.lastname: Gross
mesh_terms: D000992:Antitoxins / Q000737:chemistry* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D001427:Bacterial Toxins / Q000737:chemistry* / Q000378:metabolism*; D041961:Deuterium Exchange Measurement; D011485:Protein Binding; D017510:Protein Folding; D025941:Protein Interaction Mapping / Q000379:methods*; D013297:Streptococcus pyogenes / Q000737:chemistry* / Q000276:immunology / Q000472:pathogenicity
chemical_list: D000992:Antitoxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins