title: Using hydrogen-deuterium exchange to monitor protein structure in the presence of gold nanoparticles.
abstract: The potential applications of protein-functionalized gold nanoparticles (AuNPs) have motivated many studies characterizing protein-AuNP interactions. However, the lack of detailed structural information has hindered our ability to understand the mechanism of protein adsorption on AuNPs. In order to determine the structural perturbations that occur during adsorption, hydrogen/deuterium exchange (HDX) of amide protons was measured for two proteins by NMR. Specifically, we measured both slow (5-300 min) and fast (10-500 ms) H/D exchange rates for GB3 and ubiquitin, two well-characterized proteins. Overall, amide exchange rates are very similar in the presence and absence of AuNPs, supporting a model where the adsorbed protein remains largely folded on the AuNP surface. Small differences in exchange rates are observed for several loop residues, suggesting that the secondary structure remains relatively rigid while loops and surface residues can experience perturbations upon binding. Strikingly, several of these residues are close to lysines, which supports a model where positive surface residues may interact favorably with AuNP-bound citrate. Because these proteins appear to remain folded on AuNP surfaces, these studies suggest that it may be possible to engineer functional AuNP-based nanoconjugates without the use of chemical linkers.
authors.affiliation: Department of Chemistry, Mississippi State University , Mississippi State, Mississippi 39762, United States.
authors.forename: Ailin
authors.initials: A
authors.lastname: Wang
authors.forename: Tam
authors.initials: T
authors.lastname: Vo
authors.forename: Vu
authors.initials: V
authors.lastname: Le
authors.forename: Nicholas C
authors.initials: NC
authors.lastname: Fitzkee
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D041961:Deuterium Exchange Measurement / Q000379:methods; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D013291:Streptococcus / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D025801:Ubiquitin; D006046:Gold