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Tau missorting causes synaptic loss and neuronal dysfunction, but the mechanisms underlying both normal axonal sorting and pathological missorting remain unclear. The six human brain Tau isoforms show different axodendritic distribution, but the Tau domains governing intracellular sorting and essential interactors are unknown. Here, we aimed to identify domains or motifs of human Tau and cellular binding partners required for efficient axonal Tau sorting and to unravel isoform\u2010specific Tau interactors. Using human\n                    <jats:italic>MAPT<\/jats:italic>\n                    \u2010KO induced pluripotent stem cell (iPSC)\u2010derived glutamatergic neurons, we analyzed the sorting behavior of more than 20 truncation\u2010 or phosphorylation\u2010mutant Tau constructs and used TurboID\u2010based proximity labeling and proteomics to identify sorting\u2010 and isoform\u2010specific Tau interactors. We found that efficient axonal Tau sorting was independent of the N\u2010terminal tail, the C\u2010terminal repeat domains, AD\u2010associated phosphorylation, and the general microtubule affinity of Tau, but it requires the presence of the proline\u2010rich region 2 (PRR2). Our interactome data revealed peroxisomal accumulation of the Tau N\u2010terminal half, while axonal Tau interacted with the PP2A activator HSP110. Further, we found 0N4R\u2010specific interactions of Tau with regulators of presynaptic exocytosis and postsynaptic plasticity, which are partially associated with AD pathogenesis, including members of the CDC42 pathway and the RAB11 proteins, while 0N3R\u2010Tau bound to various cytoskeletal elements. In sum, our study i) postulates that axonal Tau sorting relies on the PRR2 domain but not on microtubule affinity and ii) unravels a potential isoform\u2010specific role in synaptic function and AD\u2010related dysfunction.\n                  <\/jats:p>","DOI":"10.1111\/acel.70215","type":"journal-article","created":{"date-parts":[[2025,9,24]],"date-time":"2025-09-24T13:37:40Z","timestamp":1758721060000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Tau Axonal Sorting and Interaction With Synaptic Plasticity Modulators Is Domain\u2010 and Isoform\u2010Dependent in Human\n                    <scp>iPSC<\/scp>\n                    \u2010Derived Neurons"],"prefix":"10.1111","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0681-901X","authenticated-orcid":false,"given":"Michael","family":"Bell\u2010Simons","sequence":"first","affiliation":[{"name":"Institute of Human Genetics University Hospital 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(CMMC) University of Cologne  Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Adam","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics University Hospital Cologne  Cologne Germany"},{"name":"Center for Molecular Medicine Cologne (CMMC) University of Cologne  Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Klimek","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics University Hospital Cologne  Cologne Germany"},{"name":"Center for Molecular Medicine Cologne (CMMC) University of Cologne  Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Buchholz","sequence":"additional","affiliation":[{"name":"Institute of Human Genetics University Hospital Cologne  Cologne Germany"},{"name":"Center for Molecular Medicine Cologne (CMMC) University of Cologne  Cologne 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