{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T17:04:42Z","timestamp":1790874282179,"version":"4.1.0"},"reference-count":49,"publisher":"The Company of Biologists","issue":"10","license":[{"start":{"date-parts":[[2000,5,15]],"date-time":"2000-05-15T00:00:00Z","timestamp":958348800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/www.biologists.com\/user-licence-1-1\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2000,5,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>Tight junctions (TJ) of polarized epithelial cells regulate barrier function at mucosal surfaces. Structural proteins of TJs include hyperphosphorylated occludin (HO) and the peripheral membrane protein, ZO-1. Since TJs are dynamically regulated, and lipid-modified signal transduction proteins localize to TJs, we considered the possibility that the TJ itself is composed of microdomains with unique structure. Differential detergent extraction and isopycnic sucrose density gradients were utilized to isolate TJ-enriched membranes from a polarized intestinal epithelial cell line, T84. Here we report that major pools of hyperphosphorylated occludin (HO) and ZO-1 are found in raft-like membrane microdomains with characteristics of the previously described detergent-insoluble glycolipid rafts (DIGs). Properties of such gradient fractions included Triton X-100 (TX-100) insolubility, light scattering at 600 nm, buoyant density of approximately 1.08 g\/cm3 and increased cholesterol content compared to high density fractions. Similar results were obtained using natural epithelium. Unlike the TJ proteins HO and ZO-1, other basolateral transmembrane proteins including E-cadherin, c-met and beta 1 integrin were not increased in DIG-like fractions. Immunoprecipitation studies revealed coprecipitation of a pool of occludin with caveolin-1, a scaffolding protein abundant in DIGs. Coprecipitation results were supported by immunofluorescence and immunogold labeling studies demonstrating caveolin-1 localization in the apical membrane and focal colocalization with occludin in TJs. TJ disassembly by calcium chelation resulted in displacement of TJ proteins from the \u2018raft-like\u2019 compartment. Our findings suggest that raft-like compartments play an important role in the spatial organization of TJs and probably in regulation of paracellular permeability in epithelial cells.<\/jats:p>","DOI":"10.1242\/jcs.113.10.1771","type":"journal-article","created":{"date-parts":[[2021,4,25]],"date-time":"2021-04-25T00:24:36Z","timestamp":1619310276000},"page":"1771-1781","source":"Crossref","is-referenced-by-count":372,"title":["Tight junctions are membrane microdomains"],"prefix":"10.1242","volume":"113","author":[{"given":"A.","family":"Nusrat","sequence":"first","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. A.","family":"Parkos","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P.","family":"Verkade","sequence":"additional","affiliation":[{"name":"2 Cell Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. S.","family":"Foley","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T. W.","family":"Liang","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W.","family":"Innis-Whitehouse","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. K.","family":"Eastburn","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. L.","family":"Madara","sequence":"additional","affiliation":[{"name":"Emory University School of Medicine 1 Epithelial Pathobiology Research Unit, Department of Pathology , , Atlanta GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2000,5,15]]},"reference":[{"key":"2024071016060684800_JOCES_113_10_1771C1","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1093\/clinchem\/20.4.470","article-title":"Enzymatic determination of total serum cholesterol","volume":"20","author":"Allain","year":"1974","journal-title":"Clin. Chem"},{"key":"2024071016060684800_JOCES_113_10_1771C2","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1083\/jcb.106.4.1141","article-title":"Characterization of ZO-1, a protein component of the tight junction from mouse liver and Madin-Darby canine kidney cells","volume":"106","author":"Anderson","year":"1988","journal-title":"J. Cell Biol"},{"key":"2024071016060684800_JOCES_113_10_1771C3","first-page":"G467","article-title":"Tight junctions and the molecular basis for regulation of paracellular permeability","volume":"269","author":"Anderson","year":"1995","journal-title":"Am. J. Physiol"},{"key":"2024071016060684800_JOCES_113_10_1771C4","doi-asserted-by":"crossref","first-page":"10909","DOI":"10.1073\/pnas.90.23.10909","article-title":"Caveolae: where incoming and outgoing messengers meet","volume":"90","author":"Anderson","year":"1993","journal-title":"Proc. Natl. Acad. Sci. 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Physiol"}],"container-title":["Journal of Cell Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.biologists.com\/jcs\/article-pdf\/113\/10\/1771\/3547935\/joces_113_10_1771.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.biologists.com\/jcs\/article-pdf\/113\/10\/1771\/3547935\/joces_113_10_1771.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,10]],"date-time":"2024-07-10T16:10:51Z","timestamp":1720627851000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/jcs\/article\/113\/10\/1771\/2659\/Tight-junctions-are-membrane-microdomains"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2000,5,15]]},"references-count":49,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2000,5,15]]}},"URL":"https:\/\/doi.org\/10.1242\/jcs.113.10.1771","relation":{},"ISSN":["0021-9533","1477-9137"],"issn-type":[{"value":"0021-9533","type":"print"},{"value":"1477-9137","type":"electronic"}],"subject":[],"published":{"date-parts":[[2000,5,15]]}}}