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Here, we describe the multi-omic chemical validation of\n                    <jats:italic>Plasmodium<\/jats:italic>\n                    M1 alanyl metalloaminopeptidase as an attractive drug target using the selective inhibitor, MIPS2673. MIPS2673 demonstrated potent inhibition of recombinant\n                    <jats:italic>Plasmodium falciparum<\/jats:italic>\n                    (\n                    <jats:italic>Pf<\/jats:italic>\n                    A-M1) and\n                    <jats:italic>Plasmodium vivax<\/jats:italic>\n                    (\n                    <jats:italic>Pv<\/jats:italic>\n                    A-M1) M1 metalloaminopeptidases, with selectivity over other\n                    <jats:italic>Plasmodium<\/jats:italic>\n                    and human aminopeptidases, and displayed excellent in vitro antimalarial activity with no significant host cytotoxicity. Orthogonal label-free chemoproteomic methods based on thermal stability and limited proteolysis of whole parasite lysates revealed that MIPS2673 solely targets\n                    <jats:italic>Pf<\/jats:italic>\n                    A-M1 in parasites, with limited proteolysis also enabling estimation of the binding site on\n                    <jats:italic>Pf<\/jats:italic>\n                    A-M1 to within ~5 \u00c5 of that determined by X-ray crystallography. Finally, functional investigation by untargeted metabolomics demonstrated that MIPS2673 inhibits the key role of\n                    <jats:italic>Pf<\/jats:italic>\n                    A-M1 in haemoglobin digestion. Combined, our unbiased multi-omic target deconvolution methods confirmed the on-target activity of MIPS2673, and validated selective inhibition of M1 alanyl metalloaminopeptidase as a promising antimalarial strategy.\n                  <\/jats:p>","DOI":"10.7554\/elife.92990","type":"journal-article","created":{"date-parts":[[2024,2,2]],"date-time":"2024-02-02T09:25:50Z","timestamp":1706865950000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.92990","source":"Crossref","is-referenced-by-count":5,"title":["Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as an antimalarial strategy"],"prefix":"10.7554","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6654-7266","authenticated-orcid":true,"given":"Carlo","family":"Giannangelo","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02bfwt286","id-type":"ROR","asserted-by":"publisher"}],"name":"Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical 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