Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/4120
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dc.contributor.authorÖzçelik, Serdar-
dc.contributor.authorOrr, Galya-
dc.contributor.authorHu, Dehong-
dc.contributor.authorChen, Chii-Shiarng-
dc.contributor.authorReşat, Haluk-
dc.contributor.authorHarms, Greg S.-
dc.contributor.authorOpresko, Lee K.-
dc.contributor.authorJohn Wiley and Sons Inc., H. Steven-
dc.contributor.authorColson, Steven D.-
dc.date.accessioned2014-07-25T03:29:49Z
dc.date.available2014-07-25T03:29:49Z
dc.date.issued2013-10-08en_US
dc.identifier.citationÖzçelik, S., Orr, G., Hu, D., Chen, C.-S., Resat, H., Harms, G.S., Opresko, L.K., Wiley, H.S., & Colson, S.D. (June 21, 2004). FRET measurements between small numbers of molecules identifies subtle changes in receptor interactions. Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, 119. doi:10.1117/12.536770en
dc.identifier.urihttp://dx.doi.org/10.1117/12.536770en_US
dc.identifier.urihttp://hdl.handle.net/11147/4120
dc.description.abstractOverexpression of HER2 alters the cellular behavior of EGF receptor (EGFR) and itself,with great implications on cell fate. To understand the molecular interactions underlying these alterations, we quantified the association between the two receptors by looking at efficiency changes in fluorescence resonance energy transfer (FRET) between a small number of molecules at the membrane of living cells. Human mammary epithelial (HME) cells expressing varying degrees of HER2 were studied, to identify and compare the degree of receptors interactions as a function of HER2 overexpression. A high resolution wide-field laser microscope combined with a high sensitivity cooled CCD camera was used to capture simultaneously donor and acceptor emissions. Alternating between green and red lasers every 80 msec, donor, FRET, and acceptor images were acquired and were used to calculate FRET efficiency. Automated image analysis was developed to create FRET efficiency maps from overlapping donor, acceptor and FRET images, and derive FRET efficiency histograms to quantify receptorreceptor interactions pixel by pixel. This approach enabled us to detect subtle changes in the average distance between EGFR molecules, and between EGFR and HER2. We found pre-existing EGFR homoassociations, and EGFR-HER2 heteroassociations in cells overexpressing HER2, and identified the changes in these interactions with ligand stimulation. These observations demonstrate the power of FRET measurements between small numbers of molecules in identifying subtle changes in molecular interactions in living cell.en
dc.language.isoenen_US
dc.publisherSPIEen_US
dc.relation.ispartofProceedings of the SPIEen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectFRETen
dc.subjectFluorescenceen
dc.subjectCellsen
dc.subjectDimerizationen
dc.subjectEGFRen
dc.subjectHER2en
dc.titleFRET measurements between small numbers of molecules identifies subtle changes in receptor interactionsen_US
dc.typeArticleen_US
dc.authoridTR7497
dc.institutionauthorÖzçelik, Serdar-
dc.departmentIzmir Institute of Technology. Chemistryen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.identifier.doi10.1117/12.536770-
dc.relation.doi10.1117/12.536770en_US
dc.coverage.doi10.1117/12.536770en_US
item.languageiso639-1en-
item.openairetypeArticle-
item.cerifentitytypePublications-
item.grantfulltextopen-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.fulltextWith Fulltext-
crisitem.author.dept04.01. Department of Chemistry-
Appears in Collections:Chemistry / Kimya
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection
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