Fig. 5

Identification of AgNPs– and Ag⁺–binding proteins based on proteomic and metallomic strategies. a A schematic showing “protocol 1” for proteomic analysis of protein adsorbed on AgNPs. Experimental procedure is presented in the upper panel. Briefly, a does of 50, 100 or 200 μg AgNPs was added into HepG2 cell lysate for 1 h incubation at 37 °C, and adsorbed proteins on AgNPs were isolated and identified using SDS-PAGE and HPLC-MS/MS. b A schematic showing “protocol 2” for metallomic analysis of Ag-protein complexes in AgNPs-treated HepG2 cells at 1, 2 or 4 μg/mL for 24 h. Experimental procedure is presented in the upper panel. Briefly, untreated or AgNPs-treated HepG2 cells were harvested for cell lysis. Then, the soluble Ag-protein complexes in cell lysate were separated and analyzed using SDS-PAGE and ICP-MS. Finally, the detected Ag⁺-binding proteins were identified using HPLC-MS/MS. c Identification of protein targets by both “protocol 1” and “protocol 2” depicted as a Venn diagram. The detailed information of these proteins was provided in Additional file 1: Table S2 and S3. The involved biological process of proteins is obtained from UniProt (https://www.uniprot.org/)