{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Wright KM"],"funding":["NHLBI","NIDDK NIH HHS","NHLBI NIH HHS","NCI NIH HHS","NIH","NIGMS NIH HHS","NIGMS"],"pagination":["105715"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10933555"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["300(3)"],"pubmed_abstract":["NEDD4L is a HECT-type E3 ligase that catalyzes the addition of ubiquitin to intracellular substrates such as the cardiac voltage-gated sodium channel, Na<sub>V</sub>1.5. The intramolecular interactions of NEDD4L regulate its enzymatic activity which is essential for proteostasis. For Na<sub>V</sub>1.5, this process is critical as alterations in Na<sup>+</sup> current is involved in cardiac diseases including arrhythmias and heart failure. In this study, we perform extensive biochemical and functional analyses that implicate the C2 domain and the first WW-linker (1,2-linker) in the autoregulatory mechanism of NEDD4L. Through in vitro and electrophysiological experiments, the NEDD4L 1,2-linker was determined to be important in substrate ubiquitination of Na<sub>V</sub>1.5. We establish the preferred sites of ubiquitination of NEDD4L to be in the second WW-linker (2,3-linker). Interestingly, NEDD4L ubiquitinates the cytoplasmic linker between the first and second transmembrane domains of the channel (DI-DII) of Na<sub>V</sub>1.5. Moreover, we design a genetically encoded modulator of Nav1.5 that achieves Na<sup>+</sup> current reduction using the NEDD4L HECT domain as cargo of a Na<sub>V</sub>1.5-binding nanobody. These investigations elucidate the mechanisms regulating the NEDD4 family and furnish a new molecular framework for understanding Na<sub>V</sub>1.5 ubiquitination."],"journal":["The Journal of biological chemistry"],"pubmed_title":["NEDD4L intramolecular interactions regulate its auto and substrate Na<sub>V</sub>1.5 ubiquitination."],"pmcid":["PMC10933555"],"funding_grant_id":["P30 CA006973","P30 DK089502","R37 GM062437","R01 CA074305","K08 CA270403","R01 HL128743","R01 GM114250","R25 GM109441","R01 GM062437"],"pubmed_authors":["Fossier L","Gabelli SB","Kim H","Srinivasan L","Jiang H","Cole PA","Boronina T","Ben-Johny M","Paul S","Xia W","Nwafor JN","Chen Z","Nathan S","Chakouri N","Wright KM","Cole RN","Post J"],"additional_accession":[]},"is_claimable":false,"name":"NEDD4L intramolecular interactions regulate its auto and substrate Na<sub>V</sub>1.5 ubiquitination.","description":"NEDD4L is a HECT-type E3 ligase that catalyzes the addition of ubiquitin to intracellular substrates such as the cardiac voltage-gated sodium channel, Na<sub>V</sub>1.5. The intramolecular interactions of NEDD4L regulate its enzymatic activity which is essential for proteostasis. For Na<sub>V</sub>1.5, this process is critical as alterations in Na<sup>+</sup> current is involved in cardiac diseases including arrhythmias and heart failure. In this study, we perform extensive biochemical and functional analyses that implicate the C2 domain and the first WW-linker (1,2-linker) in the autoregulatory mechanism of NEDD4L. Through in vitro and electrophysiological experiments, the NEDD4L 1,2-linker was determined to be important in substrate ubiquitination of Na<sub>V</sub>1.5. We establish the preferred sites of ubiquitination of NEDD4L to be in the second WW-linker (2,3-linker). Interestingly, NEDD4L ubiquitinates the cytoplasmic linker between the first and second transmembrane domains of the channel (DI-DII) of Na<sub>V</sub>1.5. Moreover, we design a genetically encoded modulator of Nav1.5 that achieves Na<sup>+</sup> current reduction using the NEDD4L HECT domain as cargo of a Na<sub>V</sub>1.5-binding nanobody. These investigations elucidate the mechanisms regulating the NEDD4 family and furnish a new molecular framework for understanding Na<sub>V</sub>1.5 ubiquitination.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Mar","modification":"2026-06-03T04:58:27.615Z","creation":"2026-05-28T03:06:23.802Z"},"accession":"S-EPMC10933555","cross_references":{"pubmed":["38309503"],"doi":["10.1016/j.jbc.2024.105715"]}}