{"doi":"10.1038/s41467-024-50462-2","title":"Versatile nanobody-based approach to image, track and reconstitute functional Neurexin-1 in vivo","abstract":"Abstract Neurexins are key adhesion proteins that coordinate extracellular and intracellular synaptic components. Nonetheless, the low abundance of these multidomain proteins has complicated any localization and structure-function studies. Here we combine an ALFA tag (AT)/nanobody (NbALFA) tool with classic genetics, cell biology and electrophysiology to examine the distribution and function of the Drosophila Nrx-1 in vivo. We generate full-length and ΔPDZ ALFA-tagged Nrx-1 variants and find that the PDZ binding motif is key to Nrx-1 surface expression. A PDZ binding motif provided in trans, via genetically encoded cytosolic NbALFA-PDZ chimera, fully restores the synaptic localization and function of Nrx ΔPDZ-AT . Using cytosolic NbALFA-mScarlet intrabody, we achieve compartment-specific detection of endogenous Nrx-1, track live Nrx-1 transport along the motor neuron axons, and demonstrate that Nrx-1 co-migrates with Rab2-positive vesicles. Our findings illustrate the versatility of the ALFA system and pave the way towards dissecting functional domains of complex proteins in vivo.","journal":"Nature Communications","year":2024,"id":463343,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9495,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":720687,"name":"Saumitra Dey Choudhury","orcid":"0000-0001-8828-4551","position":1,"is_corresponding":false},{"id":413432,"name":"Tae Hee Han","orcid":"0000-0001-6223-9437","position":2,"is_corresponding":false},{"id":518803,"name":"Tho Huu Nguyen","orcid":null,"position":3,"is_corresponding":false},{"id":413436,"name":"Peter Nguyen","orcid":"0000-0002-1419-5975","position":4,"is_corresponding":false},{"id":1055141,"name":"Felipe Opazo","orcid":"0000-0002-4968-9713","position":5,"is_corresponding":false},{"id":413441,"name":"Mihaela Serpe","orcid":"0000-0002-9205-8589","position":6,"is_corresponding":false},{"id":413433,"name":"Rosario Vicidomini","orcid":"0000-0002-5082-3238","position":0,"is_corresponding":true}],"reference_count":81,"raw_metadata":null,"created_at":"2026-07-19T02:04:28.838123Z","pmid":"39025931","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}