Proteome‐Scale Mining of Metal‐Associated Proteins of Monkeypox Virus

Metal ions are crucial for viral processes like replication, structural integrity and immune modulation, despite that the metalloproteome of Monkeypox virus (MPXV) remains largely unexplored. Monkeypox virus is a re-emerging zoonotic Orthopoxvirus with a 197 kb genome encoding over 183 proteins. Here, in this report we aimed to identify and explore the metal-associated proteome of the MPXV. Derived from employing a structure driven pipeline, followed by the functional annotation, the subcellular localization, evolutionary aspect, and structural validation with the established experimental evidences. Yielded a set of approximately 21 % high-confidence putative metal-associated proteins with a potential as metal-binding proteins. Functional annotation, Gene Ontology (GO) enrichment, and KEGG Orthology (KO) term assignment also revealed significant enrichment of pathways primarily related to functions such as, viral replication, genome maintenance, transcription, virion assembly, and host immune modulation. These metal-associated proteins are hypothesized to perform critical biological roles throughout the viral life cycle and pathogenesis. This includes nucleotide metabolism, transcriptional regulation, redox balance and immune evasion, and virion morphogenesis. These findings were further strengthened by subcellular localization analysis, which predicts the presence of metalloproteins within MPXV and its viral factories. Further, suggesting the spatial distribution of various metals and its utilization in the host organism. Facilitating the activities from viral attachment, entry, replication, transcription, viral assembly, and release as either mature virion (MV) or intracellular mature virion (IMV). Comparison and mapping these identified metal-associated MPXV proteins to Vaccinia virus followed by the virus-host network analysis highlighted the proposed role of metal-associated proteins within conserved Orthopoxvirus interaction pathways.