000878733 001__ 878733
000878733 005__ 20220930130249.0
000878733 0247_ $$2doi$$a10.1186/s12896-020-00633-y
000878733 0247_ $$2Handle$$a2128/25559
000878733 0247_ $$2pmid$$apmid:32854679
000878733 0247_ $$2WOS$$aWOS:000567175700001
000878733 037__ $$aFZJ-2020-03033
000878733 082__ $$a610
000878733 1001_ $$0P:(DE-Juel1)145762$$aGruteser, Nadine$$b0
000878733 245__ $$aEstablishing a sensitive fluorescence-based quantification method for cyclic nucleotides
000878733 260__ $$aLondon$$bBioMed Central$$c2020
000878733 3367_ $$2DRIVER$$aarticle
000878733 3367_ $$2DataCite$$aOutput Types/Journal article
000878733 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1614335428_24429
000878733 3367_ $$2BibTeX$$aARTICLE
000878733 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000878733 3367_ $$00$$2EndNote$$aJournal Article
000878733 520__ $$aBackgroundApproximately 40% of prescribed drugs exert their activity via GTP-binding protein-coupled receptors (GPCRs). Once activated, these receptors cause transient changes in the concentration of second messengers, e.g., cyclic adenosine 3′,5′-monophosphate (cAMP). Specific and efficacious genetically encoded biosensors have been developed to monitor cAMP fluctuations with high spatial and temporal resolution in living cells or tissue. A well characterized biosensor for cAMP is the Förster resonance energy transfer (FRET)-based Epac1-camps protein. Pharmacological characterization of newly developed ligands acting at GPCRs often includes numerical quantification of the second messenger amount that was produced.ResultsTo quantify cellular cAMP concentrations, we bacterially over-expressed and purified Epac1-camps and applied the purified protein in a cell-free detection assay for cAMP in a multi-well format. We found that the biosensor can detect as little as 0.15 pmol of cAMP, and that the sensitivity is not impaired by non-physiological salt concentrations or pH values. Notably, the assay tolerated desiccation and storage of the protein without affecting Epac1-camps cyclic nucleotide sensitivity.ConclusionsWe found that determination cAMP in lysates obtained from cell assays or tissue samples by purified Epac1-camps is a robust, fast, and sensitive assay suitable for routine and high throughput analyses.
000878733 536__ $$0G:(DE-HGF)POF3-552$$a552 - Engineering Cell Function (POF3-552)$$cPOF3-552$$fPOF III$$x0
000878733 588__ $$aDataset connected to CrossRef
000878733 7001_ $$0P:(DE-HGF)0$$aKohlhas, Viktoria$$b1
000878733 7001_ $$0P:(DE-Juel1)131909$$aBalfanz, Sabine$$b2$$ufzj
000878733 7001_ $$0P:(DE-Juel1)131923$$aFranzen, Arne$$b3$$ufzj
000878733 7001_ $$0P:(DE-HGF)0$$aGünther, Anne$$b4
000878733 7001_ $$0P:(DE-Juel1)128713$$aOffenhäusser, Andreas$$b5$$ufzj
000878733 7001_ $$0P:(DE-Juel1)131939$$aMüller, Frank$$b6$$ufzj
000878733 7001_ $$0P:(DE-HGF)0$$aNikolaev, Viacheslav$$b7
000878733 7001_ $$0P:(DE-HGF)0$$aLohse, Martin J.$$b8
000878733 7001_ $$0P:(DE-Juel1)131911$$aBaumann, Arnd$$b9$$eCorresponding author
000878733 773__ $$0PERI:(DE-600)2052746-9$$a10.1186/s12896-020-00633-y$$gVol. 20, no. 1, p. 47$$n1$$p47$$tBMC biotechnology$$v20$$x1472-6750$$y2020
000878733 8564_ $$uhttps://juser.fz-juelich.de/record/878733/files/SN-2020-00398-b.pdf
000878733 8564_ $$uhttps://juser.fz-juelich.de/record/878733/files/s12896-020-00633-y.pdf$$yOpenAccess
000878733 8564_ $$uhttps://juser.fz-juelich.de/record/878733/files/s12896-020-00633-y.pdf?subformat=pdfa$$xpdfa$$yOpenAccess
000878733 8767_ $$8SN-2020-00398-b$$92021-02-23$$d2021-02-26$$eAPC$$jDEAL$$lDEAL: Springer$$pBBIT-D-20-00025$$zFZJ-2020-02751, Listenpreis / Belegnr. 12001642441990€, 20% geprüft
000878733 909CO $$ooai:juser.fz-juelich.de:878733$$pVDB$$pdriver$$pOpenAPC$$popen_access$$popenaire$$popenCost$$pdnbdelivery
000878733 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131909$$aForschungszentrum Jülich$$b2$$kFZJ
000878733 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131923$$aForschungszentrum Jülich$$b3$$kFZJ
000878733 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)128713$$aForschungszentrum Jülich$$b5$$kFZJ
000878733 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131939$$aForschungszentrum Jülich$$b6$$kFZJ
000878733 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)131911$$aForschungszentrum Jülich$$b9$$kFZJ
000878733 9131_ $$0G:(DE-HGF)POF3-552$$1G:(DE-HGF)POF3-550$$2G:(DE-HGF)POF3-500$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bKey Technologies$$lBioSoft – Fundamentals for future Technologies in the fields of Soft Matter and Life Sciences$$vEngineering Cell Function$$x0
000878733 9132_ $$0G:(DE-HGF)POF4-899$$1G:(DE-HGF)POF4-890$$2G:(DE-HGF)POF4-800$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$aDE-HGF$$bProgrammungebundene Forschung$$lohne Programm$$vohne Topic$$x0
000878733 9141_ $$y2020
000878733 915__ $$0LIC:(DE-HGF)CCBY4$$2HGFVOC$$aCreative Commons Attribution CC BY 4.0
000878733 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)1190$$2StatID$$aDBCoverage$$bBiological Abstracts$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0600$$2StatID$$aDBCoverage$$bEbsco Academic Search$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bBMC BIOTECHNOL : 2018$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0501$$2StatID$$aDBCoverage$$bDOAJ Seal$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0500$$2StatID$$aDBCoverage$$bDOAJ$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0700$$2StatID$$aFees$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)9900$$2StatID$$aIF < 5$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess
000878733 915__ $$0StatID:(DE-HGF)0030$$2StatID$$aPeer Review$$bASC$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0561$$2StatID$$aArticle Processing Charges$$f2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0320$$2StatID$$aDBCoverage$$bPubMed Central$$d2020-01-16
000878733 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2020-01-16
000878733 920__ $$lyes
000878733 9201_ $$0I:(DE-Juel1)IBI-1-20200312$$kIBI-1$$lMolekular- und Zellphysiologie$$x0
000878733 9201_ $$0I:(DE-Juel1)IBI-3-20200312$$kIBI-3$$lBioelektronik$$x1
000878733 980__ $$ajournal
000878733 980__ $$aVDB
000878733 980__ $$aI:(DE-Juel1)IBI-1-20200312
000878733 980__ $$aI:(DE-Juel1)IBI-3-20200312
000878733 980__ $$aAPC
000878733 980__ $$aUNRESTRICTED
000878733 9801_ $$aAPC
000878733 9801_ $$aFullTexts