Updated project metadata. The race to combat antibiotic resistance and develop novel therapies has triggered studies on novel metal-based formulations. Silver remains a strong candidate since ancient times due to its multimodal and broad-spectrum activity against bacterial and fungal pathogens. N-heterocyclic carbene (NHC) complexes coordinate transition metals to generate a broad range of anticancer and/or antimicrobial agents with ongoing efforts being made to enhance lipophilicity and drug stability. The lead silver(I) acetate complex, 1,3-dibenzyl-4,5-diphenylimidazol-2-ylidene (NHC*) (SBC3) synthesised by the Tacke group has previously demonstrated promising growth and biofilm-inhibiting properties. As an extension of this, we examined the responses of two structurally different bacteria to SBC3 using label-free quantitative proteomic analysis. Multidrug resistant Pseudomonas aeruginosa (Gram-negative) and Staphylococcus aureus (Gram-positive) are associated with chronic wound infections and Cystic Fibrosis lung colonisation where co-infection often exacerbates disease. SBC3 increased the abundance of alginate biosynthesis, secretion system and drug detoxification proteins in P. aeruginosa whilst a multitude of pathways including anaerobic respiration, twitching motility, and ABC transport were decreased. This contrasted with affected pathways in S. aureus such as increased DNA replication/repair and cell redox homeostasis and decreased protein synthesis, lipoylation, glucose metabolism. Increased abundance of cell wall/membrane proteins were indicative of the structural damage induced by SBC3 to both cell types. These findings show the potential broad applications of SBC3 in treating Gram-positive and Gram-negative bacteria.