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Article: Effects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranes

TitleEffects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranes
Authors
Issue Date1989
Citation
Journal Of Receptor Research, 1989, v. 9 n. 2, p. 159-169 How to Cite?
AbstractHomogenates from dog cerebellum were fractionated using sucrose gradient centrifugation. The [3H]inositol 1,4,5-trisphosphate binding and the glucose 6-phosphatase activities were found to co-purify. The binding was saturable and had high affinity (B(max) = 44 pmol/mg protein, K(d) = 116 nM). Selective chemical modification was used to examine amino acid residues of the microsomal receptor that might be critical for the binding of inositol trisphosphate. Sulfhydryl reagents, p-chloromercuric-phenyl sulfonic acid, eosin 5-maleimide, N-ethyl maleimide and fluorescein 5-maleimide were found to be highly potent inhibitors of the binding with half-maximal inhibition occurring at about 20 μM, 70 μM, 1, mM, and 0.1 mM, respectively. The inhibition was specific since the presence of 10 μM of inositol trisphosphate during the reaction completely protected against the inhibition by these reagents. These results suggest that sulfhydryl group is essential for inositol trisphosphate binding to its receptor.
Persistent Identifierhttp://hdl.handle.net/10722/171528
ISSN
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYang, CMen_US
dc.contributor.authorLee, HCen_US
dc.date.accessioned2012-10-30T06:15:33Z-
dc.date.available2012-10-30T06:15:33Z-
dc.date.issued1989en_US
dc.identifier.citationJournal Of Receptor Research, 1989, v. 9 n. 2, p. 159-169en_US
dc.identifier.issn0197-5110en_US
dc.identifier.urihttp://hdl.handle.net/10722/171528-
dc.description.abstractHomogenates from dog cerebellum were fractionated using sucrose gradient centrifugation. The [3H]inositol 1,4,5-trisphosphate binding and the glucose 6-phosphatase activities were found to co-purify. The binding was saturable and had high affinity (B(max) = 44 pmol/mg protein, K(d) = 116 nM). Selective chemical modification was used to examine amino acid residues of the microsomal receptor that might be critical for the binding of inositol trisphosphate. Sulfhydryl reagents, p-chloromercuric-phenyl sulfonic acid, eosin 5-maleimide, N-ethyl maleimide and fluorescein 5-maleimide were found to be highly potent inhibitors of the binding with half-maximal inhibition occurring at about 20 μM, 70 μM, 1, mM, and 0.1 mM, respectively. The inhibition was specific since the presence of 10 μM of inositol trisphosphate during the reaction completely protected against the inhibition by these reagents. These results suggest that sulfhydryl group is essential for inositol trisphosphate binding to its receptor.en_US
dc.languageengen_US
dc.relation.ispartofJournal of Receptor Researchen_US
dc.subject.meshAnimalsen_US
dc.subject.meshBinding Sitesen_US
dc.subject.meshCalcium Channelsen_US
dc.subject.meshCerebellum - Metabolismen_US
dc.subject.meshDogsen_US
dc.subject.meshInositol 1,4,5-Trisphosphateen_US
dc.subject.meshInositol 1,4,5-Trisphosphate Receptorsen_US
dc.subject.meshInositol Phosphates - Metabolismen_US
dc.subject.meshReceptors, Cell Surface - Analysisen_US
dc.subject.meshReceptors, Cytoplasmic And Nuclearen_US
dc.subject.meshSugar Phosphates - Metabolismen_US
dc.subject.meshSulfhydryl Reagents - Pharmacologyen_US
dc.titleEffects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranesen_US
dc.typeArticleen_US
dc.identifier.emailLee, HC:leehc@hku.hken_US
dc.identifier.authorityLee, HC=rp00545en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.3109/10799898909066051-
dc.identifier.pmid2545875-
dc.identifier.scopuseid_2-s2.0-0024356511en_US
dc.identifier.volume9en_US
dc.identifier.issue2en_US
dc.identifier.spage159en_US
dc.identifier.epage169en_US
dc.identifier.isiWOS:A1989AA44200004-
dc.identifier.scopusauthoridYang, CM=37025138700en_US
dc.identifier.scopusauthoridLee, HC=26642959100en_US

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