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Article: Na +-H + exchanger 3 (NHE3) is present in lipid rafts in the rabbit ileal brush border: A role for rafts in trafficking and rapid stimulation of NHE3

TitleNa +-H + exchanger 3 (NHE3) is present in lipid rafts in the rabbit ileal brush border: A role for rafts in trafficking and rapid stimulation of NHE3
Authors
Issue Date2001
PublisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0022-3751
Citation
Journal Of Physiology, 2001, v. 537 n. 2, p. 537-552 How to Cite?
Abstract1. Rabbit ileal Na +-absorbing cell Na +-H + exchanger 3 (NHE3) was shown to exist in three pools in the brush border (BB), including a population in lipid rafts. Approximately 50% of BB NHE3 was associated with Triton X-100-soluble fractions and the other ∼50% with Triton X-100-insoluble fractions; ∼33% of the detergent-insoluble NHE3 was present in cholesterol-enriched lipid microdomains (rafts). 2. The raft pool of NHE3 was involved in the stimulation of BB NHE3 activity with epidermal growth factor (EGF). Both EGF and clonidine treatments were associated with a rapid increase in the total amount of BB NHE3. This EGF- and clonidine-induced increase of BB NHE3 was associated with an increase in the raft pool of NHE3 and to a smaller extent with an increase in the total detergent-insoluble fraction, but there was no change in the detergent-soluble pool. In agreement with the rapid increase in the amount of NHE3 in the BB, EGF also caused a rapid stimulation of BB Na +-H + exchange activity. 3. Disrupting rafts by removal of cholesterol with methyl-β-cyclodextrin (MβCD) or destabilizing the actin cytoskeleton with cytochalasin D decreased the amount of NHE3 in early endosomes isolated by OptiPrep gradient fractionation. Specifically, NHE3 was shown to associate with endosomal vesicles immunoisolated by anti-EEA1 (early endosomal autoantigen 1) antibodycoated magnetic beads and the endosome-associated NHE3 was decreased by cytochalasin D and MβCD treatment. 4. We conclude that: (i) a pool of ileal BB NHE3 exists in lipid rafts; (ii) EGF and clonidine increase the amount of BB NHE3; (iii) lipid rafts and to a lesser extent, the cytoskeleton, but not the detergent-soluble NHE3 pool, are involved in the EGF- and clonidine-induced acute increase in amount of BB NHE3; (iv) lipid rafts and the actin cytoskeleton play important roles in the basal endocytosis of BB NHE3.
Persistent Identifierhttp://hdl.handle.net/10722/171271
ISSN
2015 Impact Factor: 4.731
2015 SCImago Journal Rankings: 2.670
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLi, Xen_US
dc.contributor.authorGalli, Ten_US
dc.contributor.authorWade, JBen_US
dc.contributor.authorWade, JBen_US
dc.contributor.authorWeinman, EJen_US
dc.contributor.authorLeung, Gen_US
dc.contributor.authorCheong, Aen_US
dc.contributor.authorLouvard, Den_US
dc.contributor.authorDonowitz, Men_US
dc.date.accessioned2012-10-30T06:13:05Z-
dc.date.available2012-10-30T06:13:05Z-
dc.date.issued2001en_US
dc.identifier.citationJournal Of Physiology, 2001, v. 537 n. 2, p. 537-552en_US
dc.identifier.issn0022-3751en_US
dc.identifier.urihttp://hdl.handle.net/10722/171271-
dc.description.abstract1. Rabbit ileal Na +-absorbing cell Na +-H + exchanger 3 (NHE3) was shown to exist in three pools in the brush border (BB), including a population in lipid rafts. Approximately 50% of BB NHE3 was associated with Triton X-100-soluble fractions and the other ∼50% with Triton X-100-insoluble fractions; ∼33% of the detergent-insoluble NHE3 was present in cholesterol-enriched lipid microdomains (rafts). 2. The raft pool of NHE3 was involved in the stimulation of BB NHE3 activity with epidermal growth factor (EGF). Both EGF and clonidine treatments were associated with a rapid increase in the total amount of BB NHE3. This EGF- and clonidine-induced increase of BB NHE3 was associated with an increase in the raft pool of NHE3 and to a smaller extent with an increase in the total detergent-insoluble fraction, but there was no change in the detergent-soluble pool. In agreement with the rapid increase in the amount of NHE3 in the BB, EGF also caused a rapid stimulation of BB Na +-H + exchange activity. 3. Disrupting rafts by removal of cholesterol with methyl-β-cyclodextrin (MβCD) or destabilizing the actin cytoskeleton with cytochalasin D decreased the amount of NHE3 in early endosomes isolated by OptiPrep gradient fractionation. Specifically, NHE3 was shown to associate with endosomal vesicles immunoisolated by anti-EEA1 (early endosomal autoantigen 1) antibodycoated magnetic beads and the endosome-associated NHE3 was decreased by cytochalasin D and MβCD treatment. 4. We conclude that: (i) a pool of ileal BB NHE3 exists in lipid rafts; (ii) EGF and clonidine increase the amount of BB NHE3; (iii) lipid rafts and to a lesser extent, the cytoskeleton, but not the detergent-soluble NHE3 pool, are involved in the EGF- and clonidine-induced acute increase in amount of BB NHE3; (iv) lipid rafts and the actin cytoskeleton play important roles in the basal endocytosis of BB NHE3.en_US
dc.languageengen_US
dc.publisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=0022-3751en_US
dc.relation.ispartofJournal of Physiologyen_US
dc.subject.meshActins - Physiologyen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCytoskeletal Proteins - Physiologyen_US
dc.subject.meshCytoskeleton - Metabolism - Physiologyen_US
dc.subject.meshDetergentsen_US
dc.subject.meshEndocytosis - Physiologyen_US
dc.subject.meshIleum - Metabolismen_US
dc.subject.meshLipid Metabolismen_US
dc.subject.meshMaleen_US
dc.subject.meshMembrane Proteins - Metabolismen_US
dc.subject.meshMicrovilli - Metabolismen_US
dc.subject.meshPhosphoproteins - Physiologyen_US
dc.subject.meshQa-Snare Proteinsen_US
dc.subject.meshRabbitsen_US
dc.subject.meshSodium-Hydrogen Antiporter - Metabolismen_US
dc.subject.meshSolubilityen_US
dc.titleNa +-H + exchanger 3 (NHE3) is present in lipid rafts in the rabbit ileal brush border: A role for rafts in trafficking and rapid stimulation of NHE3en_US
dc.typeArticleen_US
dc.identifier.emailLeung, G:gphleung@hkucc.hku.hken_US
dc.identifier.authorityLeung, G=rp00234en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1111/j.1469-7793.2001.00537.xen_US
dc.identifier.pmid11731584-
dc.identifier.scopuseid_2-s2.0-0035576776en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035576776&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume537en_US
dc.identifier.issue2en_US
dc.identifier.spage537en_US
dc.identifier.epage552en_US
dc.identifier.isiWOS:000172624700020-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridLi, X=7501701559en_US
dc.identifier.scopusauthoridGalli, T=7006546912en_US
dc.identifier.scopusauthoridWade, JB=36153885300en_US
dc.identifier.scopusauthoridWade, JB=7201786600en_US
dc.identifier.scopusauthoridWeinman, EJ=7006627157en_US
dc.identifier.scopusauthoridLeung, G=35963668200en_US
dc.identifier.scopusauthoridCheong, A=7003847644en_US
dc.identifier.scopusauthoridLouvard, D=7005231353en_US
dc.identifier.scopusauthoridDonowitz, M=24486735400en_US

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