{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T09:56:48Z","timestamp":1753869408996,"version":"3.41.2"},"reference-count":52,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,1,2]],"date-time":"2025-01-02T00:00:00Z","timestamp":1735776000000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal Cellular Physiology"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Megalin is a multiple\u2010ligand receptor that contributes to protein reabsorption in the kidney. Recently, megalin was found to act as a novel endocytic receptor for prorenin. Internalization depended on the (pro)renin receptor. This receptor is an accessory protein of vacuolar H<jats:sup>+<\/jats:sup>\u2010ATPase (V\u2010ATPase), a complex consisting of 14 subunits and two accessory proteins. Here we explored whether V\u2010ATPase elements other than the (P)RR affect megalin\u2010mediated prorenin uptake. Using RNAi technology, we inhibited each individual V\u2010ATPase subunit in megalin\u2010expressing BN16 cells. Subsequently, we quantified megalin expression and the uptake of prorenin. To unravel the underlying molecular mechanisms, we investigated the adaptor proteins autosomal recessive hypercholesterolemia (ARH) and Disabled\u20102 (Dab2), which are important for the endocytosis of megalin, glycogen synthase kinase 3\u03b2 (GSK3\u03b2), a regulatory factor of megalin recycling, and endoplasmic reticulum stress factors (ERSF). Silencing subunit <jats:italic>Atp6v<\/jats:italic><jats:sub><jats:italic>o<\/jats:italic><\/jats:sub><jats:italic>a1<\/jats:italic> reduced prorenin uptake by 19%, while silencing accessory protein <jats:italic>Atp6ap1<\/jats:italic> increased it by 15%. Silencing other subunits exerted a more modest or no effect. Silencing <jats:italic>Atp6v<\/jats:italic><jats:sub><jats:italic>o<\/jats:italic><\/jats:sub><jats:italic>a1<\/jats:italic> reduced surface megalin density, without altering its mRNA and protein levels, and this was associated with increased GSK3\u03b2 phosphorylation and no change in ARH, Dab2, and ERSF. Silencing <jats:italic>Atp6ap1<\/jats:italic> increased megalin mRNA and protein expression and this was accompanied by upregulation of ARH and ERSF, while Dab2 expression was unaltered. In conclusion, V\u2010ATPase units differently affect megalin\u2010mediated reabsorption of prorenin, thereby offering novel pharmacological targets to not only affect renal renin\u2010angiotensin system activity, but also to treat renal diseases that are associated with disturbed protein reabsorption, like Dent's disease.<\/jats:p>","DOI":"10.1002\/jcp.31518","type":"journal-article","created":{"date-parts":[[2025,1,2]],"date-time":"2025-01-02T11:49:41Z","timestamp":1735818581000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Vacuolar H<sup>+<\/sup>\u2010ATPase and Megalin\u2010Mediated Prorenin Uptake: Focus on Elements Beyond the (Pro)Renin Receptor"],"prefix":"10.1002","volume":"240","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-2949-1398","authenticated-orcid":false,"given":"Na","family":"Wang","sequence":"first","affiliation":[{"name":"Division of Vascular Medicine and Pharmacology, Department of Internal Medicine Erasmus MC Rotterdam The Netherlands"},{"name":"Clinical Research Center, the First Affiliated Hospital of Shantou University Medical College Shantou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xifeng","family":"Lu","sequence":"additional","affiliation":[{"name":"Clinical Research Center, the First Affiliated Hospital of Shantou University Medical College Shantou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. 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