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001-es BibID:	BIBFORM042481
035-os BibID:	(scopus)40049093067 (wos)000253576200006
Első szerző:	Fazekas Zsolt (biofizikus)
Cím:	Two-sided fluorescence resonance energy transfer for assessing molecular interactions of up to three distinct species in confocal microscopy / Zsolt Fazekas, Miklós Petrás, Ákos Fábián, Zsuzsanna Pályi-Krekk, Péter Nagy, Sándor Damjanovich, György Vereb, János Szöllősi
Dátum:	2008
ISSN:	1552-4922 1552-4930
Megjegyzések:	The role of the expression patterns of proteins involved in oncogenesis can be understood after characterizing their multimolecular interactions. Conventional FRET methods permit the analysis of interaction between two molecular species at the most, which necessitates the introduction of new approaches for studying multicomponent signaling complexes. Flow cytometric as well as microscopic donor (dbFRET) and acceptor (abFRET) photobleaching FRET measurements were performed to determine the association states of ErbB2, b1-integrin, and CD44 receptors. Based on consecutively applied abFRET and dbFRET methods (two-sided FRET), the relationship of b1-integrin?ErbB2 heteroassociation to ErbB2 homoassociation and of b1-integrin?ErbB2 heteroassociation to ErbB2?CD44 heteroassociation was studied by correlating pixel-by-pixel FRET values of the corresponding abFRET and dbFRET images in contour plots. Anticorrelation was observed between b1-integrin?ErbB2 heteroassociation and ErbB2 homoassociation on trastuzumab sensitive N87 and SK-BR-3 cells, while modest positive correlation was found between b1-integrin?ErbB2 and ErbB2?CD44 heteroassociationon trastuzumab resistant MKN-7 cells. The FRET efficiency values of b1-integrin?ErbB2 heteroassociation were markedly higher at the focal adhesion regions on attachedcells than those measured by flow cytometry on detached cells. In conclusion, we implemented an experimental set-up termed two-sided FRET for correlating two pairwiseinteractions of three arbitrarily chosen molecular species. On the basis of our results, we assume that the homoassociation state of ErbB2 is dynamically modulated by its interaction with b1-integrins.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk beta1-integrin abFRET analysis Biophysics Carcinoma CD44 Cell Adhesion Cell Adhesion Molecules Cell Line Cells Cholera Toxin Confocal Confocal microscopy cytology dbFRET Energy Transfer ErbB2 Flow Cytometry Fluorescein Fluorescence fluorescence microscopy Fluorescence Resonance Energy Transfer FRET Hiv-1 Human Hungary Image Cytometry Image Processing Kinetics Laser scanning confocal microscopy Luminescence methods Microscopy Photobleaching Protein-protein interactions Proteins Receptor tyrosine kinase Research Signal Transduction Software therapy Trastuzumab resistance tsFRET Tyrosine egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry Part A. - 73 : 3 (2008), p. 209-219. -
További szerzők:	Petrás Miklós (1977-) (orvos) Fábián Ákos István (1982-) (aneszteziológus) Pályiné Krekk Zsuzsanna (1974-) (molekuláris biológus) Nagy Péter (1971-) (biofizikus) Damjanovich Sándor (1936-2017) (biofizikus) Vereb György (1965-) (biofizikus, orvos) Szöllősi János (1953-) (biofizikus)
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001-es BibID:	BIBFORM004886
Első szerző:	Szentesi Gergely (kémia-fizika tanár)
Cím:	Computer program for analyzing donor photobleaching FRET image series / Szentesi, G., Vereb, G., Horvath, G., Bodnar, A., Fabian, A., Matko, J., Gaspar, R., Damjanovich, S., Matyus, L., Jenei, A.
Dátum:	2005
ISSN:	1552-4922
Megjegyzések:	The photobleaching fluorescence resonance energy transfer (pbFRET) technique is a spectroscopic method to measure proximity relations between fluorescently labeled macromolecules using digital imaging microscopy. To calculate the energy transfer values one has to determine the bleaching time constants in pixel-by-pixel fashion from the image series recorded on the donor-only and donor and acceptor double-labeled samples. Because of the large number of pixels and the time-consuming calculations, this procedure should be assisted by powerful image data processing software. There is no commercially available software that is able to fulfill these requirements. METHODS: New evaluation software was developed to analyze pbFRET data for Windows platform in National Instrument LabVIEW 6.1. This development environment contains a mathematical virtual instrument package, in which the Levenberg-Marquardt routine is also included. As a reference experiment, FRET efficiency between the two chains (beta2-microglobulin and heavy chain) of major histocompatibility complex (MHC) class I glycoproteins and FRET between MHC I and MHC II molecules were determined in the plasma membrane of JY, human B lymphoma cells. RESULTS: The bleaching time constants calculated on pixel-by-pixel basis can be displayed as a color-coded map or as a histogram from raw image format. CONCLUSION: In this report we introduce a new version of pbFRET analysis and data processing software that is able to generate a full analysis pattern of donor photobleaching image series under various conditions.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Algorithms analysis beta 2-Microglobulin Biophysics Cell Line,Tumor Cells Energy Transfer Fluorescence Fluorescence Resonance Energy Transfer Glycoproteins Histocompatibility Antigens Human Humans Hungary Lymphoma Major Histocompatibility Complex metabolism methods Microscopy Photobleaching Research Software Support egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A. - 67 : 2 (2005), p. 119-128. -
További szerzők:	Vereb György (1965-) (biofizikus, orvos) Horváth Gábor (1974-) (biofizikus) Dóczy-Bodnár Andrea (1970-) (biofizikus) Fábián Ákos István (1982-) (aneszteziológus) Matkó János (1952-) (biológus) Gáspár Rezső (1944-) (biofizikus) Damjanovich Sándor (1936-2017) (biofizikus) Mátyus László (1956-) (biofizikus) Jenei Attila (1966-) (biofizikus)
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