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In kinetoplastid parasites, acidocalcisomes and their polyphosphate content are essential for osmoregulation and environmental adaptation during host switching. In this organelle, polyphosphate is synthesised and transported to the lumen by the vacuolar transporter chaperone (VTC) complex. Interestingly, unlike yeast VTC, which has five components, only two have been observed in kinetoplastids: Vtc1, which contains only a transmembrane domain and Vtc4, which, in addition to a transmembrane domain, also consists of SPX and catalytic domains. In this study, we used proximity-dependent biotinylation (BioID) in\n                    <jats:italic>Leishmania tarentolae<\/jats:italic>\n                    to identify proteins located close to the VTC complex. The complex was found near several known acidocalcisomal proteins, including membrane-bound pyrophosphatase (mPPase), vacuolar-type H\u2009\u207a\u2009-ATPase (V-H\n                    <jats:sup>+<\/jats:sup>\n                    -ATPase), Ca\u00b2\u2009\u207a\u2009-transporting P-type ATPase (Ca\n                    <jats:sup>2+<\/jats:sup>\n                    -ATPase), zinc transporter (ZnT), and palmitoyl acyltransferase 2 (PAT2). Importantly, this approach revealed three novel VTC binding partners (VBPs) that colocalise and interact with the complex in acidocalcisomes, as confirmed by confocal microscopy, pulldown assays, and AlphaFold3 structural predictions. Together, our results expand the acidocalcisome interactome and suggest that the newly identified VBPs may contribute to the structural organisation and regulatory function of the VTC complex in phosphate homeostasis of kinetoplastid parasites.\n                  <\/jats:p>","DOI":"10.1371\/journal.pntd.0014511","type":"journal-article","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T17:36:59Z","timestamp":1783964219000},"page":"e0014511","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Novel binding partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania 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