000001302 001__ 1302
000001302 005__ 20200610184538.0
000001302 0247_ $$2Handle$$a2128/13952
000001302 037__ $$aPreJuSER-1302
000001302 0881_ $$aJuel-4272
000001302 088__ $$2JUEL$$aJuel-4272
000001302 1001_ $$0P:(DE-Juel1)VDB57288$$avon der Hocht, Iris$$b0$$eCoresponding author$$uFZJ
000001302 245__ $$aFluorescence Spectroscopy of Recoverin Function and Conformation
000001302 260__ $$aJülich$$bForschungszentrum Jülich GmbH Zentralbibliothek, Verlag$$c2008
000001302 300__ $$a119 p.
000001302 3367_ $$0PUB:(DE-HGF)11$$2PUB:(DE-HGF)$$aDissertation / PhD Thesis
000001302 3367_ $$0PUB:(DE-HGF)3$$2PUB:(DE-HGF)$$aBook
000001302 3367_ $$02$$2EndNote$$aThesis
000001302 3367_ $$2DRIVER$$adoctoralThesis
000001302 3367_ $$2BibTeX$$aPHDTHESIS
000001302 3367_ $$2DataCite$$aOutput Types/Dissertation
000001302 3367_ $$2ORCID$$aDISSERTATION
000001302 4900_ $$0PERI:(DE-600)2414853-2$$832540$$aBerichte des Forschungszentrums Jülich$$v4272$$x0944-2952
000001302 502__ $$aKöln, Univ., Diss., 2008$$bDr. (Univ.)$$cUniv. Köln$$d2008
000001302 500__ $$aRecord converted from VDB: 12.11.2012
000001302 520__ $$aRecoverin is a neuronal calcium sensor protein (NCS-protein) from the vertebrate photoreceptor which is involved in light adaptation. Recoverin changes its conformation upon sequential binding of two calcium ions. This conformational change induces the extrusion of a covalently attached myristoyl residue from a hydrophobic binding pocket which enables Recoverin to interact with lipid membranes (calcium-myristoyl switch). In this thesis, I report on my investigation of Recoverin’s calcium myristoyl switch, using a recently developed modification of fluorescence correlation spectroscopy (FCS) that is called dual-focus FCS (2fFCS). This method allows for measuring absolute diffusion coefficients with an accuracy of better than a few percent. Recoverin, and the Recoverin mutants E85Q and E121Q which bind only one or no Ca$^{2+}$, respectively, were labeled with the fluorescent dye Alexa647. Differences in the hydrodynamic radius due to conformational changes of Recoverin and its mutants upon calcium binding were monitored by measuring the diffusion coefficient of these molecules as a function of free calcium concentration. The calcium dependent interaction of Recoverin with lipid membranes was measured in solutions of small unilamellar vesicles (SUVs) of different lipid composition. Again, diffusion measurements were used to determine the fraction of free and lipid-bound Recoverin using the strongly different diffusion coefficients of both fractions. To account for the fluorescence brightness difference of the dye label when in solution and close to a lipid membrane, a new three-photon correlation analysis was developed and tested.
000001302 536__ $$0G:(DE-Juel1)FUEK412$$2G:(DE-HGF)$$aGrundlagen für zukünftige Informationstechnologien$$cP42$$x0
000001302 655_7 $$aHochschulschrift$$xDissertation (Univ.)
000001302 8564_ $$uhttps://juser.fz-juelich.de/record/1302/files/J%C3%BCl_4272_Hocht.pdf$$yOpenAccess
000001302 8564_ $$uhttps://juser.fz-juelich.de/record/1302/files/J%C3%BCl_4272_Hocht.pdf?subformat=pdfa$$xpdfa$$yOpenAccess
000001302 909CO $$ooai:juser.fz-juelich.de:1302$$pdnbdelivery$$pVDB$$pdriver$$popen_access$$popenaire
000001302 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess
000001302 9141_ $$y2008
000001302 9131_ $$0G:(DE-Juel1)FUEK412$$bSchlüsseltechnologien$$kP42$$lGrundlagen für zukünftige Informationstechnologien (FIT)$$vGrundlagen für zukünftige Informationstechnologien$$x0
000001302 9201_ $$0I:(DE-Juel1)VDB804$$d31.12.2008$$gINB$$kINB-1$$lZelluläre Biophysik$$x1
000001302 970__ $$aVDB:(DE-Juel1)102313
000001302 9801_ $$aFullTexts
000001302 980__ $$aVDB
000001302 980__ $$aConvertedRecord
000001302 980__ $$aphd
000001302 980__ $$aI:(DE-Juel1)ICS-4-20110106
000001302 980__ $$aUNRESTRICTED
000001302 981__ $$aI:(DE-Juel1)IBI-1-20200312
000001302 981__ $$aI:(DE-Juel1)ICS-4-20110106