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Here, we report the X\u2010ray crystal structure of\n                    <jats:italic>Pf<\/jats:italic>\n                    HAM1, which revealed a homodimeric structure, additionally validated by size\u2010exclusion chromatography\u2013multi\u2010angle light scattering analysis. The two monomeric units in the dimer were aligned in a parallel fashion, and critical residues associated with substrate and metal binding were identified, wherein a notable structural difference was observed in the \u03b2\u2010sheet main frame compared to human inosine triphosphate pyrophosphatase.\n                    <jats:italic>Pf<\/jats:italic>\n                    HAM1 exhibited Mg\n                    <jats:sup>++<\/jats:sup>\n                    \u2010dependent pyrophosphohydrolase activity and the highest binding affinity to dITP compared to other non\u2010canonical nucleotides as measured by isothermal titration calorimetry. Modifying the\n                    <jats:italic>pfham1<\/jats:italic>\n                    genomic locus followed by live\u2010cell imaging of expressed mNeonGreen\u2010tagged\n                    <jats:italic>Pf<\/jats:italic>\n                    HAM1 demonstrated its ubiquitous presence in the cytoplasm across erythrocytic stages with greater expression in trophozoites and schizonts. Interestingly, CRISPR\u2010Cas9\/DiCre recombinase\u2010guided\n                    <jats:italic>pfham1<\/jats:italic>\n                    \u2010null\n                    <jats:italic>P. falciparum<\/jats:italic>\n                    survived in culture under standard growth conditions, indicating its assistive role in non\u2010canonical nucleotide clearance during intra\u2010erythrocytic stages. This is the first comprehensive structural and functional report of\n                    <jats:italic>Pf<\/jats:italic>\n                    HAM1, an atypical nucleotide\u2010cleansing enzyme in\n                    <jats:italic>P.\u2009falciparum.<\/jats:italic>\n                  <\/jats:p>","DOI":"10.1111\/febs.17216","type":"journal-article","created":{"date-parts":[[2024,7,14]],"date-time":"2024-07-14T10:06:07Z","timestamp":1720951567000},"page":"4349-4371","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Structure\u2013function analysis of nucleotide housekeeping protein\n                    <scp>HAM1<\/scp>\n                    from human malaria parasite\n                    <i>Plasmodium falciparum<\/i>"],"prefix":"10.1111","volume":"291","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8311-530X","authenticated-orcid":false,"given":"Debanjan","family":"Saha","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases and Immunology CSIR\u2010Indian Institute of Chemical Biology  Kolkata India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4967-6963","authenticated-orcid":false,"given":"Atanu","family":"Pramanik","sequence":"additional","affiliation":[{"name":"Division of Structural Biology &amp; Bioinformatics CSIR\u2010Indian Institute of Chemical Biology  Kolkata India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0982-5688","authenticated-orcid":false,"given":"Aline","family":"Freville","sequence":"additional","affiliation":[{"name":"Department of Infection Biology London School of Hygiene &amp; Tropical Medicine  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