{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,27]],"date-time":"2025-11-27T02:59:05Z","timestamp":1764212345740,"version":"3.41.2"},"reference-count":45,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2024,3,27]],"date-time":"2024-03-27T00:00:00Z","timestamp":1711497600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Acta Physiologica"],"published-print":{"date-parts":[[2024,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Aim<\/jats:title><jats:p>Trafficking, membrane retention, and signal\u2010specific regulation of the Na<jats:sup>+<\/jats:sup>\/H<jats:sup>+<\/jats:sup> exchanger 3 (NHE3) are modulated by the Na<jats:sup>+<\/jats:sup>\/H<jats:sup>+<\/jats:sup> Exchanger Regulatory Factor (NHERF) family of PDZ\u2010adapter proteins. This study explored the assembly of NHE3 and NHERF2 with the cGMP\u2010dependent kinase II (cGKII) within detergent\u2010resistant membrane microdomains (DRMs, \u201clipid rafts\u201d) during in vivo guanylate cycle C receptor (Gucy2c) activation in murine small intestine.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Small intestinal brush border membranes (<jats:italic>siBBM<\/jats:italic>s) were isolated from wild type, NHE3\u2010deficient, cGMP\u2010kinase II\u2010deficient, and NHERF2\u2010deficient mice, after oral application of the heat\u2010stable <jats:italic>Escherichia coli<\/jats:italic> toxin (STa) analog linaclotide. Lipid raft and non\u2010raft fractions were separated by Optiprep density gradient centrifugation of Triton X\u2010solubilized <jats:italic>siBBM<\/jats:italic>s. Confocal microscopy was performed to study NHE3 redistribution after linaclotide application in vivo.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>In the WT <jats:italic>siBBM<\/jats:italic>, NHE3, NHERF2, and cGKII were strongly raft associated. The raft association of NHE3, but not of cGKII, was NHERF2 dependent. After linaclotide application to WT mice, lipid raft association of NHE3 decreased, that of cGKII increased, while that of NHERF2 did not change. NHE3 expression in the BBM shifted from a microvillar to a terminal web region. The linaclotide\u2010induced decrease in NHE3 raft association and in microvillar abundance was abolished in cGKII\u2010deficient mice, and strongly reduced in NHERF2\u2010deficient mice.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>NHE3, cGKII, and NHERF2 form a lipid raft\u2010associated signal complex in the <jats:italic>siBBM<\/jats:italic>, which mediates the inhibition of salt and water absorption by Gucy2c activation. NHERF2 enhances the raft association of NHE3, which is essential for its close interaction with the exclusively raft\u2010associated activated cGKII.<\/jats:p><\/jats:sec>","DOI":"10.1111\/apha.14125","type":"journal-article","created":{"date-parts":[[2024,3,27]],"date-time":"2024-03-27T12:01:58Z","timestamp":1711540918000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["<scp>cGMP<\/scp>\u2010dependent kinase 2, Na<sup>+<\/sup>\/H<sup>+<\/sup> exchanger <scp>NHE3,<\/scp> and <scp>PDZ<\/scp>\u2010adaptor <scp>NHERF2<\/scp> co\u2010assemble in apical membrane microdomains"],"prefix":"10.1111","volume":"240","author":[{"given":"Min","family":"Luo","sequence":"first","affiliation":[{"name":"Department of Gastroenterology, Hepatology, Infectiology and Endocrinology Hannover Medical School  Hannover Germany"},{"name":"Department of Infectious Diseases The Second Affiliated Hospital of Chongqing Medical University  Chongqing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongjian","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology, Infectiology and Endocrinology Hannover Medical School  Hannover Germany"},{"name":"Department of Endocrinology The Second Affiliated Hospital of Chongqing Medical University  Chongqing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6534-9831","authenticated-orcid":false,"given":"Katerina","family":"Nikolovska","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology, Infectiology and Endocrinology Hannover Medical School  Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brigitte","family":"Riederer","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology, Infectiology and Endocrinology Hannover Medical School  Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Enrico","family":"Patrucco","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Pharmakologie und Toxikologie TU M\u00fcnchen  M\u00fcnchen Germany"},{"name":"Department of Molecular Biotechnology and Health Science University of Torino  Torino Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Franz","family":"Hofmann","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Pharmakologie und Toxikologie TU M\u00fcnchen  M\u00fcnchen Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9600-2769","authenticated-orcid":false,"given":"Ursula","family":"Seidler","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology, Infectiology and Endocrinology Hannover Medical School  Hannover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,3,27]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2022.899286"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrgastro.2015.111"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ph.55.030193.003215"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.medmal.2020.10.009"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pharmthera.2004.08.007"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI119683"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1997.7487"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiv300"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2015-309441"},{"key":"e_1_2_9_11_1","doi-asserted-by":"crossref","DOI":"10.14309\/ctg.0000000000000427","article-title":"A potential treatment of congenital sodium diarrhea in patients with activating GUCY2C mutations","volume":"12","author":"Vugt AHM","year":"2021","journal-title":"Clin Transl Gastroenterol"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1097\/MOG.0000000000000020"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2018.00299"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5295.2082"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0016-5085(00)70419-7"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00135.2006"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M704878200"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2020.114040"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M500505200"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2010.198721"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-008-0579-1"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1159\/000185553"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E14-12-1597"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-007-0239-x"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2010.356"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M601740200"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e16-12-0862"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1159\/000356577"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-291X(02)00412-6"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1177\/36.7.3290330"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-7793.2001.00537.x"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2010.03.015"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1517\/17425255.2011.572875"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.12.7025"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.00160.2004"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00526.2008"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.40.25856"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.470799"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00191.2010"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/969"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2007.131722"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00193.2003"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00261.2017"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1111\/apha.13498"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23179993"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2009.09.055"}],"container-title":["Acta Physiologica"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/apha.14125","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T09:48:03Z","timestamp":1715075283000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/apha.14125"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,27]]},"references-count":45,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2024,4]]}},"alternative-id":["10.1111\/apha.14125"],"URL":"https:\/\/doi.org\/10.1111\/apha.14125","archive":["Portico"],"relation":{},"ISSN":["1748-1708","1748-1716"],"issn-type":[{"type":"print","value":"1748-1708"},{"type":"electronic","value":"1748-1716"}],"subject":[],"published":{"date-parts":[[2024,3,27]]},"assertion":[{"value":"2023-10-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-02-14","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-03-27","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e14125"}}