Development of a dual reporter system to simultaneously visualize ca2+ signals and ampk activity

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dc.authorid0000-0003-0352-1947
dc.authorid0000-0002-9373-0808
dc.contributor.authorErdoğan, Yusuf Ceyhun
dc.contributor.authorPilic, Johannes
dc.contributor.authorGottschalk, Benjamin
dc.contributor.authorYiğit, Esra Nur
dc.contributor.authorZaki, Asal Ghaffari
dc.contributor.authorÖztürk, Gürkan
dc.contributor.authorEroğlu, Emrah
dc.contributor.authorOkutan, Begüm
dc.contributor.authorSommer, Nicole G.
dc.contributor.authorWeinberg, Annelie M.
dc.contributor.authorSchindl, Rainer
dc.contributor.authorGraier, Wolfgang F.
dc.contributor.authorMalli, Roland
dc.date.accessioned2024-11-04T08:02:12Z
dc.date.available2024-11-04T08:02:12Z
dc.date.issued2024
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Rejeneratif ve Restoratif Tıp Araştırmaları Merkezi (REMER)
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Sağlık Bilim ve Teknolojileri Araştırma Enstitüsü
dc.departmentİstanbul Medipol Üniversitesi, Uluslararası Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Fizyoloji Ana Bilim Dalı
dc.description.abstractIn this study, we introduce a new separation of phases-based activity reporter of kinase (SPARK) for AMP-activated kinase (AMPK), named AMPK-SPARK, which reports the AMPK activation by forming bright fluorescent clusters. Furthermore, we introduce a dual reporter system, named GCaMP-AMPK-SPARK, by incorporating a single-fluorescent protein (FP)-based Ca2+ biosensor, GCaMP6f, into our initial design, enabling simultaneous monitoring of Ca2+ levels and AMPK activity. This system offers the essential quality of information by single-channel fluorescence microscopy without the need for coexpression of different biosensors and elaborate filter layouts to overcome spectral limitations. We used AMPK-SPARK to map endogenous AMPK activity in different cell types and visualized the dynamics of AMPK activation in response to various stimuli. Using GCaMP-AMPK-SPARK, we revealed cell-to-cell heterogeneities in AMPK activation by Ca2+ mobilization. We anticipate that this dual reporter strategy can be employed to study the intricate interplays between different signaling networks and kinase activities.
dc.description.sponsorshipMedical University of Graz ; Nikon Austria within the Nikon-Center of Excellence, Graz ; Austrian Science Fund (FWF) ; Metabolic and Cardiovascular Disease ; Austrian infrastructure program 2013/2014, Nikon Austria Inc. ; BioTechMed, Grazen_US
dc.identifier.citationErdoğan, Y. C., Pilic, J., Gottschalk, B., Yiğit, E. N., Zaki, A. G., Öztürk, G. ... Malli, R. (2024). Development of a dual reporter system to simultaneously visualize ca2+ signals and ampk activity. ACS Sensors, 9(9), 4680-4689. http://dx.doi.org/10.1021/acssensors.4c01058
dc.identifier.doi10.1021/acssensors.4c01058
dc.identifier.endpage4689
dc.identifier.issn2379-3694
dc.identifier.issue9
dc.identifier.pmid39167044
dc.identifier.scopus2-s2.0-85201763901
dc.identifier.scopusqualityQ1
dc.identifier.startpage4680
dc.identifier.urihttp://dx.doi.org/10.1021/acssensors.4c01058
dc.identifier.urihttps://hdl.handle.net/20.500.12511/12845
dc.identifier.volume9
dc.identifier.wos001295999300001en_US
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorYiğit, Esra Nur
dc.institutionauthorZaki, Asal Ghaffari
dc.institutionauthorÖztürk, Gürkan
dc.institutionauthorEroğlu, Emrah
dc.language.isoen
dc.relation.ecinfo:eu-repo/grantAgreement/EC/FP7/I3716-B27
dc.relation.ecinfo:eu-repo/grantAgreement/EC/FP7/I5474
dc.relation.ecinfo:eu-repo/grantAgreement/EC/FP7/DK-W1226
dc.relation.ecinfo:eu-repo/grantAgreement/EC/FP7/P27070
dc.relation.ispartofACS Sensorsen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsAttribution 4.0 International*
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/*
dc.subjectAMPK Reporter
dc.subjectCa2+ Biosensor
dc.subjectFluorescence Microscopy
dc.subjectGcamp
dc.subjectMultiplexing
dc.subjectPhase Separation-Based Activity Reporter Of Kinase
dc.subjectSPARK
dc.titleDevelopment of a dual reporter system to simultaneously visualize ca2+ signals and ampk activity
dc.typeArticle

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