000056498 001__ 56498
000056498 005__ 20200402205905.0
000056498 0247_ $$2pmid$$apmid:17078090
000056498 0247_ $$2DOI$$a10.1002/prot.21231
000056498 0247_ $$2WOS$$aWOS:000243358000018
000056498 037__ $$aPreJuSER-56498
000056498 041__ $$aeng
000056498 082__ $$a540
000056498 084__ $$2WoS$$aBiochemistry & Molecular Biology
000056498 084__ $$2WoS$$aBiophysics
000056498 1001_ $$0P:(DE-Juel1)131924$$aGensch, T.$$b0$$uFZJ
000056498 245__ $$aCa2+-dependent conformational changes in the neuronal Ca2+-sensor recoverin probed by the fluorescent dye Alexa647
000056498 260__ $$aNew York, NY$$bWiley-Liss$$c2007
000056498 300__ $$a492 - 499
000056498 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article
000056498 3367_ $$2DataCite$$aOutput Types/Journal article
000056498 3367_ $$00$$2EndNote$$aJournal Article
000056498 3367_ $$2BibTeX$$aARTICLE
000056498 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000056498 3367_ $$2DRIVER$$aarticle
000056498 440_0 $$016361$$aProteins - Structure Function and Bioinformatics$$v66$$x0887-3585
000056498 500__ $$aRecord converted from VDB: 12.11.2012
000056498 520__ $$aRecoverin belongs to the superfamily of EF-hand Ca2+-binding proteins and operates as a Ca2+-sensor in vertebrate photoreceptor cells, where it regulates the activity of rhodopsin kinase GRK1 in a Ca2+-dependent manner. Ca2+-dependent conformational changes in recoverin are allosterically controlled by the covalently attached myristoyl group. The amino acid sequence of recoverin harbors a unique cysteine at position 38. The cysteine can be modified by the fluorescent dye Alexa647 using a maleimide-thiol coupling step. Introduction of Alexa647 into recoverin did not disturb the biological function of recoverin, as it can regulate rhodopsin kinase activity like unlabeled recoverin. Performance of the Ca2+-myristoyl switch of labeled recoverin was monitored by Ca2+-dependent association with immobilized lipids using surface plasmon resonance spectroscopy. When the Ca2+-concentration was varied, labeled myristoylated recoverin showed a 37%-change in fluorescence emission and a 34%-change in excitation intensity, emission and excitation maxima shifted by 6 and 18 nm, respectively. In contrast, labeled nonmyristoylated recoverin exhibited only minimal changes. Time-resolved fluorescence measurements showed biexponentiell fluorescence decay, in which the slower time constant of 2 ns was specifically influenced by Ca2+-induced conformational changes. A similar influence on the slower time constant was observed with the recoverin mutant RecE85Q that has a disabled EF-hand 2, but no such influence was detected with the mutant RecE121Q (EF-hand 3 is nonfunctional) that contains the myristoyl group in a clamped position. We conclude from our results that Alexa647 bound to cysteine 38 can monitor the conformational transition in recoverin that is under control of the myristoyl group.
000056498 536__ $$0G:(DE-Juel1)FUEK409$$2G:(DE-HGF)$$aFunktion und Dysfunktion des Nervensystems$$cP33$$x0
000056498 588__ $$aDataset connected to Web of Science, Pubmed
000056498 650_2 $$2MeSH$$aAmino Acid Substitution
000056498 650_2 $$2MeSH$$aAnimals
000056498 650_2 $$2MeSH$$aCalcium: pharmacology
000056498 650_2 $$2MeSH$$aCalcium: physiology
000056498 650_2 $$2MeSH$$aCattle
000056498 650_2 $$2MeSH$$aCyclic AMP: analogs & derivatives
000056498 650_2 $$2MeSH$$aCyclic AMP: chemistry
000056498 650_2 $$2MeSH$$aCysteine: chemistry
000056498 650_2 $$2MeSH$$aFluorescent Dyes: chemistry
000056498 650_2 $$2MeSH$$aG-Protein-Coupled Receptor Kinase 1: metabolism
000056498 650_2 $$2MeSH$$aModels, Molecular
000056498 650_2 $$2MeSH$$aMutagenesis, Site-Directed
000056498 650_2 $$2MeSH$$aMutation, Missense
000056498 650_2 $$2MeSH$$aMyristic Acid: chemistry
000056498 650_2 $$2MeSH$$aPoint Mutation
000056498 650_2 $$2MeSH$$aProtein Binding
000056498 650_2 $$2MeSH$$aProtein Conformation
000056498 650_2 $$2MeSH$$aProtein Processing, Post-Translational
000056498 650_2 $$2MeSH$$aProtein Structure, Tertiary
000056498 650_2 $$2MeSH$$aRecombinant Fusion Proteins: chemistry
000056498 650_2 $$2MeSH$$aRecoverin: chemistry
000056498 650_2 $$2MeSH$$aRecoverin: drug effects
000056498 650_2 $$2MeSH$$aRecoverin: genetics
000056498 650_2 $$2MeSH$$aSpectrometry, Fluorescence
000056498 650_2 $$2MeSH$$aStructure-Activity Relationship
000056498 650_2 $$2MeSH$$aSurface Plasmon Resonance
000056498 650_7 $$00$$2NLM Chemicals$$aFluorescent Dyes
000056498 650_7 $$00$$2NLM Chemicals$$aRCV1 protein, Bos taurus
000056498 650_7 $$00$$2NLM Chemicals$$aRecombinant Fusion Proteins
000056498 650_7 $$0135844-11-0$$2NLM Chemicals$$aRecoverin
000056498 650_7 $$039824-30-1$$2NLM Chemicals$$a8-aminohexylamino cAMP
000056498 650_7 $$052-90-4$$2NLM Chemicals$$aCysteine
000056498 650_7 $$0544-63-8$$2NLM Chemicals$$aMyristic Acid
000056498 650_7 $$060-92-4$$2NLM Chemicals$$aCyclic AMP
000056498 650_7 $$07440-70-2$$2NLM Chemicals$$aCalcium
000056498 650_7 $$0EC 2.7.11.14$$2NLM Chemicals$$aG-Protein-Coupled Receptor Kinase 1
000056498 650_7 $$2WoSType$$aJ
000056498 65320 $$2Author$$aEF-hand
000056498 65320 $$2Author$$aCa2+-binding protein
000056498 65320 $$2Author$$amyristoyl switch
000056498 65320 $$2Author$$afluorescence emission and excitation
000056498 7001_ $$0P:(DE-HGF)0$$aKomolov, K. E.$$b1
000056498 7001_ $$0P:(DE-HGF)0$$aSenin, I. I.$$b2
000056498 7001_ $$0P:(DE-HGF)0$$aPhilippov, P. P.$$b3
000056498 7001_ $$0P:(DE-HGF)0$$aKoch, K.-W.$$b4
000056498 773__ $$0PERI:(DE-600)1475032-6$$a10.1002/prot.21231$$gVol. 66, p. 492 - 499$$p492 - 499$$q66<492 - 499$$tProteins$$v66$$x0887-3585$$y2007
000056498 8567_ $$uhttp://dx.doi.org/10.1002/prot.21231
000056498 909CO $$ooai:juser.fz-juelich.de:56498$$pVDB
000056498 9131_ $$0G:(DE-Juel1)FUEK409$$bGesundheit$$kP33$$lFunktion und Dysfunktion des Nervensystems$$vFunktion und Dysfunktion des Nervensystems$$x0
000056498 9141_ $$y2007
000056498 915__ $$0StatID:(DE-HGF)0010$$aJCR/ISI refereed
000056498 9201_ $$0I:(DE-Juel1)VDB804$$d31.12.2008$$gINB$$kINB-1$$lZelluläre Biophysik$$x0
000056498 970__ $$aVDB:(DE-Juel1)88673
000056498 980__ $$aVDB
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000056498 980__ $$aI:(DE-Juel1)ICS-4-20110106
000056498 980__ $$aUNRESTRICTED
000056498 981__ $$aI:(DE-Juel1)IBI-1-20200312
000056498 981__ $$aI:(DE-Juel1)ICS-4-20110106