The nano-bio interface mapped by oxidative footprinting of the adsorption sites of myoglobin

Oxidative footprinting has been used to study the structure of macromolecular assemblies such as protein-protein and protein-ligand complexes. We propose a novel development of this technique to probe the protein corona that forms at the surface of nanoparticles in any biological medium. Indeed, very few techniques allow studying this interface at the molecular and residue level. Based on hydroxyl radical-mediated oxidation of proteins and analysis by nanoscale liquid chromatography coupled to tandem mass spectrometry (nanoLC-MS/MS), two sites of adsorption of myoglobin on silica nanoparticles are identified. This method gives new insights in the understanding of protein adsorption on nanomaterials. Graphical Abstract The adsorption sites of myoglobin on silica nanoparticles are mapped by oxidative footprinting of the free (a) and adsorbed (b) protein