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Differentiating between inactive and active states of rhodopsin by atomic force microscopy in native membranes

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cris.lastimport.scopus2024-02-12T19:42:55Z
dc.abstract.enMembrane proteins, including G protein-coupled receptors (GPCRs), present a challenge in studying their structural properties under physiological conditions. Moreover, to better understand the activity of proteins requires examination of single molecule behaviors rather than ensemble averaged behaviors. Force–distance curve-based AFM (FD-AFM) was utilized to directly probe and localize the conformational states of a GPCR within the membrane at nanoscale resolution based on the mechanical properties of the receptor. FD-AFM was applied to rhodopsin, the light receptor and a prototypical GPCR, embedded in native rod outer segment disc membranes from photoreceptor cells of the retina in mice. Both FD-AFM and computational studies on coarse-grained models of rhodopsin revealed that the active state of the receptor has a higher Young’s modulus compared to the inactive state of the receptor. Thus, the inactive and active states of rhodopsin could be differentiated based on the stiffness of the receptor. Differentiating the states based on the Young’s modulus allowed for the mapping of the different states within the membrane. Quantifying the active states present in the membrane containing the constitutively active G90D rhodopsin mutant or apoprotein opsin revealed that most receptors adopt an active state. Traditionally, constitutive activity of GPCRs has been described in terms of two-state models where the receptor can achieve only a single active state. FD-AFM data are inconsistent with a two-state model but instead require models that incorporate multiple active states.
dc.affiliationUniwersytet Warszawski
dc.contributor.authorCieplak, Marek
dc.contributor.authorFilipek, Sławomir
dc.contributor.authorPark, Paul
dc.contributor.authorSenapati, Subhadip
dc.contributor.authorBernaola, Adolfo Poma
dc.date.accessioned2024-01-24T21:49:00Z
dc.date.available2024-01-24T21:49:00Z
dc.date.copyright2019-05-10
dc.date.issued2019
dc.description.accesstimeAT_PUBLICATION
dc.description.financeNie dotyczy
dc.description.number11
dc.description.versionFINAL_PUBLISHED
dc.description.volume91
dc.identifier.doi10.1021/ACS.ANALCHEM.9B00546
dc.identifier.issn0003-2700
dc.identifier.urihttps://repozytorium.uw.edu.pl//handle/item/104872
dc.languageeng
dc.pbn.affiliationchemical sciences
dc.relation.ispartofAnalytical Chemistry
dc.relation.pages7226–7235
dc.rightsCC-BY
dc.sciencecloudnosend
dc.subject.enYoung’s modulus
dc.subject.enRodent models
dc.subject.enPeptides and proteins
dc.subject.enReceptors
dc.subject.enMembranes
dc.titleDifferentiating between inactive and active states of rhodopsin by atomic force microscopy in native membranes
dc.typeJournalArticle
dspace.entity.typePublication