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Article: Aspartic acid scanning mutation analysis of a goldfish growth hormone-releasing hormone (GHRH) receptor specific to the GHRHsalmon-like peptide

TitleAspartic acid scanning mutation analysis of a goldfish growth hormone-releasing hormone (GHRH) receptor specific to the GHRHsalmon-like peptide
Authors
KeywordsGHRH
GHRH receptor
GHRHcatfish-like
GHRHsalmon-like
Scanning mutation
Issue Date2005
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygcen
Citation
General And Comparative Endocrinology, 2005, v. 140 n. 1, p. 41-51 How to Cite?
AbstractGrowth hormone-releasing hormone (GHRH) plays a pivotal role in the regulation of growth. The study of goldfish GHRH and its receptor is of particular interest as it is so far the only animal model in which two forms of GHRH-like (catfish-like and salmon-like) peptides coexist, and these peptides share only 30-40% of amino acid sequence identities with their mammalian counterparts. For these reasons, we have previously characterized a goldfish GHRH receptor, which is specific for a synthetic carp GHRH-like peptide. In this study, we investigated the structure-function relationships between the receptor and various ligands. Interestingly, among the two endogenous goldfish GHRH-like peptides, only the GHRHsalmon-like peptide was able to stimulate CHO cells transfected with the goldfish GHRH receptor. When the receptor was challenged by GHRHsalmon-like peptide either continuously for 45 min or periodically at 45-min intervals, mild homologous desensitization was observed. To determine whether the negatively charged residues of the receptor are responsible for discriminating GHRHsalmon-like from GHRHcatfish-like, 10 aspartic acid residues residing in the N-terminal ectodomain and the second exoloop were individually mutated to alanine by site-directed mutagenesis. Among these 10 mutants, four of them (D66A, D122A, D 190A, and D196A) were defective as indicated by both cAMP assays and extracellular acidification rate measurements. Confocal microscopic studies showed that the D66A and D122A mutants, but not the D190A and D196A mutants, were expressed properly at the plasma membrane. Collectively, these results suggest that aspartic acid residues at positions 66 and 122 are critical for the interaction between the goldfish GHRH receptor and its endogenous ligands. © 2004 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/69579
ISSN
2015 Impact Factor: 2.667
2015 SCImago Journal Rankings: 1.245
ISI Accession Number ID
References
Grants

 

DC FieldValueLanguage
dc.contributor.authorKee, Fen_HK
dc.contributor.authorNg, SSMen_HK
dc.contributor.authorVaudry, Hen_HK
dc.contributor.authorPang, RTKen_HK
dc.contributor.authorLau, EHYen_HK
dc.contributor.authorChan, SMen_HK
dc.contributor.authorChow, BKCen_HK
dc.date.accessioned2010-09-06T06:14:58Z-
dc.date.available2010-09-06T06:14:58Z-
dc.date.issued2005en_HK
dc.identifier.citationGeneral And Comparative Endocrinology, 2005, v. 140 n. 1, p. 41-51en_HK
dc.identifier.issn0016-6480en_HK
dc.identifier.urihttp://hdl.handle.net/10722/69579-
dc.description.abstractGrowth hormone-releasing hormone (GHRH) plays a pivotal role in the regulation of growth. The study of goldfish GHRH and its receptor is of particular interest as it is so far the only animal model in which two forms of GHRH-like (catfish-like and salmon-like) peptides coexist, and these peptides share only 30-40% of amino acid sequence identities with their mammalian counterparts. For these reasons, we have previously characterized a goldfish GHRH receptor, which is specific for a synthetic carp GHRH-like peptide. In this study, we investigated the structure-function relationships between the receptor and various ligands. Interestingly, among the two endogenous goldfish GHRH-like peptides, only the GHRHsalmon-like peptide was able to stimulate CHO cells transfected with the goldfish GHRH receptor. When the receptor was challenged by GHRHsalmon-like peptide either continuously for 45 min or periodically at 45-min intervals, mild homologous desensitization was observed. To determine whether the negatively charged residues of the receptor are responsible for discriminating GHRHsalmon-like from GHRHcatfish-like, 10 aspartic acid residues residing in the N-terminal ectodomain and the second exoloop were individually mutated to alanine by site-directed mutagenesis. Among these 10 mutants, four of them (D66A, D122A, D 190A, and D196A) were defective as indicated by both cAMP assays and extracellular acidification rate measurements. Confocal microscopic studies showed that the D66A and D122A mutants, but not the D190A and D196A mutants, were expressed properly at the plasma membrane. Collectively, these results suggest that aspartic acid residues at positions 66 and 122 are critical for the interaction between the goldfish GHRH receptor and its endogenous ligands. © 2004 Elsevier Inc. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ygcenen_HK
dc.relation.ispartofGeneral and Comparative Endocrinologyen_HK
dc.subjectGHRHen_HK
dc.subjectGHRH receptoren_HK
dc.subjectGHRHcatfish-likeen_HK
dc.subjectGHRHsalmon-likeen_HK
dc.subjectScanning mutationen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshAspartic Aciden_HK
dc.subject.meshCHO Cellsen_HK
dc.subject.meshCell Culture Techniquesen_HK
dc.subject.meshCricetinaeen_HK
dc.subject.meshCricetulusen_HK
dc.subject.meshCyclic AMP - analysisen_HK
dc.subject.meshDNA Mutational Analysisen_HK
dc.subject.meshGoldfish - physiologyen_HK
dc.subject.meshGrowth Hormone-Releasing Hormone - genetics - pharmacologyen_HK
dc.subject.meshLigandsen_HK
dc.subject.meshModels, Animalen_HK
dc.subject.meshReceptors, Neuropeptide - genetics - physiologyen_HK
dc.subject.meshReceptors, Pituitary Hormone-Regulating Hormone - genetics - physiologyen_HK
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_HK
dc.subject.meshSalmonen_HK
dc.subject.meshStructure-Activity Relationshipen_HK
dc.subject.meshTransfectionen_HK
dc.titleAspartic acid scanning mutation analysis of a goldfish growth hormone-releasing hormone (GHRH) receptor specific to the GHRHsalmon-like peptideen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0016-6480&volume=140&issue=1&spage=41&epage=51&date=2005&atitle=Aspartic+acid+scanning+mutation+analysis+of+a+goldfish+growth+hormone-releasing+hormone+(GHRH)+receptor+specific+to+the+GHRHsalmon-like+peptide.en_HK
dc.identifier.emailNg, SSM: ssmng@hku.hken_HK
dc.identifier.emailPang, RTK: rtkpang@hku.hken_HK
dc.identifier.emailChow, BKC: bkcc@hku.hken_HK
dc.identifier.authorityNg, SSM=rp00767en_HK
dc.identifier.authorityPang, RTK=rp01761en_HK
dc.identifier.authorityChow, BKC=rp00681en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.ygcen.2004.09.010en_HK
dc.identifier.pmid15596070-
dc.identifier.scopuseid_2-s2.0-21644484167en_HK
dc.identifier.hkuros114255en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-21644484167&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume140en_HK
dc.identifier.issue1en_HK
dc.identifier.spage41en_HK
dc.identifier.epage51en_HK
dc.identifier.isiWOS:000226170800005-
dc.publisher.placeUnited Statesen_HK
dc.relation.projectFunctional and structural evolution of VIP and PACAP receptors in vertebrates-
dc.relation.projectThe concerted hypophysiotropic actions of GHRH and PACAP on the control of growth hormone release in goldfish, Carassius auratus-
dc.identifier.scopusauthoridKee, F=8973454200en_HK
dc.identifier.scopusauthoridNg, SSM=7403358718en_HK
dc.identifier.scopusauthoridVaudry, H=35446602600en_HK
dc.identifier.scopusauthoridPang, RTK=7004376636en_HK
dc.identifier.scopusauthoridChan, SM=7404255669en_HK
dc.identifier.scopusauthoridChow, BKC=7102826193en_HK

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