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1.

001-es BibID:BIBFORM057536
Első szerző:Bene László (biofizikus)
Cím:À la Fizeau in flow : pulse shape-assisted fluorescence lifetime / László Bene, János Szöllősi
Dátum:2014
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok hozzászólás
folyóiratcikk
fluorescence
resonance
energy
transfer
Megjelenés:Cytometry Part A. - 85 : 12 (2014), p. 991-994. -
További szerzők:Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:OTKA Bridging Fund
OTKA
OSTRAT/810/213 University of Debrecen
Egyéb
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
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2.

001-es BibID:BIBFORM006027
Első szerző:Bene László (biofizikus)
Cím:Lateral organization of the ICAM-1 molecule at the surface of human lymphoblasts : a possible model for its co-distribution with the IL-2 receptor, class I and class II HLA molecules / Bene L., Balázs M., Matkó J., Möst J., Dierich M. P., Szöllösi J., Damjanovich S.
Dátum:1994
Megjegyzések:Lateral distribution of the ICAM-1 molecule and its topological relationship (mutual proximity) to the heavy and light chains of class I HLA molecules, HLA-DR and interleukin-2 receptor alpha-chain (IL-2R alpha) were studied in the plasma membrane of HUT-102B2 T and JY B lymphoblastoid cell lines by the technique of flow cytometric energy transfer (FCET). Effects of adherency and treatments with recombinant interferon-gamma or tumor necrosis factor-alpha on the relative expression level of ICAM-1 to the above cell surface proteins were also investigated. While the cytokines did not significantly affect the ICAM-1 level of either cell line, an increased ICAM-1 expression was found on adherent JY cells. The ICAM-1 expression varied significantly with the cell cycle and culture conditions, as well. The statistical analysis of the differences observed in the energy transfer efficiency histograms resulted in a possible model of lateral co-distribution of these proteins in the plasma membrane. These two-dimensional patterns proved to be different for T and B lymphoma lines. ICAM-1 molecules showed a high degree of self-association on HUT-102B2 (T) cells, while they were mainly expressed as monomers on the surface of JY (B) cells. Both cells showed a significant (ca. 30%) difference between densities of the heavy and light chains of class I HLA antigen, suggesting a substantial amount of beta 2-microglobulin free heavy chains on these cell lines. The class I HLA molecules also showed partial self-association, but on both cell lines. The beta 2-microglobulin and the heavy chain of the class I HLA showed strongly different proximities to the IL-2R alpha, HLA-DR and ICAM-1 molecules, indicating that their orientations relative to the other proteins are dissimilar. IL-2R alpha molecules of the HUT-102B2 (T) cells are located mostly in the vicinity of the beta 2-microglobulin. In contrast, the local density of HLA-DR antigens is higher in the proximity of the heavy chain than in the vicinity of the beta 2-microglobulin. The possible functional significance of these protein patterns is also discussed herein.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
analysis
Antibodies,Monoclonal
B-Lymphocyte Subsets
beta 2-Microglobulin
Cell Adhesion
Cell Adhesion Molecules
Cell Cycle
Cell Line
Energy Transfer
Flow Cytometry
Histocompatibility Antigens Class I
HLA Antigens
HLA-D Antigens
HLA-DR Antigens
Human
Hungary
immunology
Intercellular Adhesion Molecule-1
Interferon Type II
Interleukin-2
Light
physiology
Receptors,Interleukin-2
Support,Non-U.S.Gov't
Support,U.S.Gov't,Non-P.H.S.
T-Lymphocyte Subsets
Tumor Necrosis Factor
Megjelenés:European Journal of Immunology. - 24 : 9 (1994), p. 2115-2123. -
További szerzők:Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Matkó János (1952-) (biológus) Most, J. Dierich, Manfred P. Szöllősi János (1953-) (biofizikus) Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:DOI
Borító:

3.

001-es BibID:BIBFORM004858
Első szerző:Bene László (biofizikus)
Cím:Detection of receptor trimers on the cell surface by flow cytometric fluorescence energy homotransfer measurements / László Bene, János Szöllősi, Gergely Szentesi, László Damjanovich, Rezső Gaspar, Thomas A. Waldmann, Sándor Damjanovich
Dátum:2005
ISSN:0006-3002
Megjegyzések:Fluorescence energy homotransfer offers a powerful tool for the investigation of the state of oligomerization of cell surface receptors on a cell-by-cell basis by measuring the polarized components of fluorescence intensity of cells labeled with fluorescently stained antibodies. Here we describe homotransfer-based methods for the flow cytometric detection and analysis of hetero- and homo-associations of cell surface receptors. Homotransfer efficiencies for two- and three-body energy transfer interactions are defined and their frequency distribution curves are computed from the fluorescence anisotropy distributions of multiple-labeled cells. The fractions of receptors involved in homo-clustering is calculated based on the dependence of the fluorescence anisotropy on the surface concentration of the fluorescently stained antibodies. A homotransfer analysis of the homo- and hetero-clustering of the MHCI and MHCII glycoproteins, the cytokine receptor IL-2Ralpha, transferrin receptor and the receptor-type tyrosine phosphatase CD45 on JY B and Kit-225-K6 T cells is presented. We investigated how various factors such as the type of dye, rotational mobility of the dye and dye-targeting antibody, as well as the wavelength of the exciting light affect the homotransfer. We show that the homotransfer technique combined with the high statistical resolution of flow cytometry is an effective tool for detecting different oligomeric states of receptors by using fluorophores having restricted rotational mobility on the time scale of fluorescence
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Fluorescence anisotropy
Receptor clustering
MHCI and MHCII glycoprotein
IL-2Rα
Transferrin receptor
CD45
Megjelenés:Biochimica et Biophysica Acta (BBA). Molecular Cell Research. - 1744 : 2 (2005), p. 176-198. -
További szerzők:Szöllősi János (1953-) (biofizikus) Szentesi Gergely (1976-) (kémia-fizika tanár) Damjanovich László (1960-) (általános sebész) Gáspár Rezső (1944-) (biofizikus) Waldmann, Thomas A. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:Intézményi repozitóriumban (DEA) tárolt változat
DOI
Borító:

4.

001-es BibID:BIBFORM004939
Első szerző:Bene László (biofizikus)
Cím:Major histocompatibility complex class I protein conformation altered by transmembrane potential changes / Bene, L., Szollosi, J., Balazs, M., Matyus, L., Gaspar, R., Ameloot, M., Dale, R. E., Damjanovich, S.
Dátum:1997
ISSN:0196-4763
Megjegyzések:The nature of charge distributions in membrane-bound macromolecular structures renders them susceptible to interaction with transmembrane potential fields. As a result, conformational changes in such species may be expected to occur when this potential is altered. We have detected reversible conformational change in the major histocompatibility complex (MHC) class I antigen in the plasma membrane of human JY cells, as monitored by flow-cytometric resonance energy-transfer, upon reduction of the transmembrane potential (depolarization). This change increased the intramolecular energy-transfer efficiency between fluorescent donor- and acceptor-labeled monoclonal antibodies directed, respectively, to epitopes on the light (beta 2-microglobulin) and the heavy chains of the MHC class I antigen. Repolarization of the depolarized samples restored the energy-transfer efficiency to the original values measured before depolarization. Depolarization caused similar relative changes in fluorescence resonance energy-transfer efficiency when Fab fragments were used for labeling MHC class I complex, suggesting that the observed phenomenon is not restricted to whole monoclonal antibodies.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Antibodies, Monoclonal
Antigen Presentation
B-Lymphocytes
beta 2-Microglobulin
Cell Membrane
chemistry
cytology
Dyes
Energy Transfer
Enzyme Activation
Flow Cytometry
Fluorescein-5-isothiocyanate
Fluorescence
Fluorescent Dyes
Histocompatibility Antigens Class I
Human
Hungary
immunology
Light
Major Histocompatibility Complex
Membrane Potentials
metabolism
methods
Na(+)-K(+)-Exchanging ATPase
Patch-Clamp Techniques
physiology
Protein Conformation
Rhodamines
Surface Properties
Megjelenés:Cytometry. - 27 : 4 (1997), p. 353-357. -
További szerzők:Szöllősi János (1953-) (biofizikus) Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Mátyus László (1956-) (biofizikus) Gáspár Rezső (1944-) (biofizikus) Ameloot, Marcel Dale, Robert E. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:elektronikus változat
Borító:

5.

001-es BibID:BIBFORM004703
035-os BibID:(scopus)0037013728 (wos)000176059200014
Első szerző:Damjanovich Sándor (biofizikus)
Cím:Does mosaicism of the plasma membrane at molecular and higher hierarchical levels in human lymphocytes carry information on the immediate history of cells? / Damjanovich, S., Matyus, L., Damjanovich, L., Bene, L., Jenei, A., Matko, J., Gaspar, R., Szollosi, J.
Dátum:2002
Megjegyzések:A theoretical analysis of experimental data is presented in this mini-review on non-random homo- and hetero-associations of cell surface receptors, which can be recruited in the plasma membrane or at the surface of the rough endoplasmic reticulum during the protein synthesis. In the latter case, the likely genetic origin of these supramolecular formations is analyzed, contrasting this concept to the mobility of the cell surface proteins. A model is offered which, on the one hand, allows the mobility in a restricted way even among microdomain-confined receptor proteins through 'swapping partners'. On the other hand, the lack of mixing molecular components of protein clusters will be analyzed, when homo-and hetero-associations are studied through cell fusion experiments. The most frequently studied cell surface patterns have included lipid raft organized HLA class I and II, ICAM-1, tetraspan molecules, IL2 and IL15 and other receptors, as well. On the contrary coated pit-associated transferrin receptors would not mix with the above lipid raft associated receptor patterns, although transferrin receptor would readily oligomerize into homo-associates. The functional consequences of these superstructures are also analyzed. On the 30th anniversary of the Singer-Nicolson fluid mosaic membrane model one has to pay tribute to the authors, because of their deep insight emphasizing also the mosaicism of the membranes in general and that of the plasma membrane, in particular.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
analysis
Biophysics
Cell Fusion
Cells
Human
Hungary
Lymphocytes
Proteins
egyetemen (Magyarországon) készült közlemény
Megjelenés:Immunology Letters. - 82 : 1-2 (2002), p. 93-99. -
További szerzők:Mátyus László (1956-) (biofizikus) Damjanovich László (1960-) (általános sebész) Bene László (1963-) (biofizikus) Jenei Attila (1966-) (biofizikus) Matkó János (1952-) (biológus) Gáspár Rezső (1944-) (biofizikus) Szöllősi János (1953-) (biofizikus)
Internet cím:DOI
elektronikus változat
Borító:

6.

001-es BibID:BIBFORM004645
035-os BibID:(scopus)0033552675 (wos)000084290400003
Első szerző:Damjanovich Sándor (biofizikus)
Cím:Two-dimensional receptor patterns in the plasma membrane of cells : a critical evaluation of their identification, origin and information content / Damjanovich, S., Bene, L., Matko, J., Matyus, L., Krasznai, Z., Szabo, G., Pieri, C., Gaspar, R., Szollosi, J.
Dátum:1999
Megjegyzések:A concise review is presented on the nature, possible origin and functional significance of cell surface receptor patterns in the plasma membrane of lymphoid cells. A special emphasize has been laid on the available methodological approaches, their individual virtues and sources of errors. Fluorescence energy transfer is one of the oldest available means for studying non-randomized co-distribution patterns of cell surface receptors. A detailed and critical description is given on the generation of two-dimensional cell surface receptor patterns based on pair-wise energy transfer measurements. A second hierarchical-level of receptor clusters have been described by electron and scanning force microscopies after immuno-gold-labeling of distinct receptor kinds. The origin of these receptor islands at a nanometer scale and island groups at a higher hierarchical (mum) level, has been explained mostly by detergent insoluble glycolipid-enriched complexes known as rafts, or detergent insoluble glycolipids (DIGs). These rafts are the most-likely organizational forces behind at least some kind of receptor clustering [K. Simons et al., Nature 387 (1997) 569]. These models, which have great significance in trans-membrane signaling and intra-membrane and intracellular trafficking, are accentuating the necessity to revisit the Singer-Nicolson fluid mosaic membrane model and substitute the free protein diffusion with a restricted diffusion concept.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Biophysics
Cells
Diffusion
Energy Transfer
Fluorescence
Hungary
Microscopy
Megjelenés:Biophysical Chemistry. - 82 : 2-3 (1999), p. 99-108. -
További szerzők:Bene László (1963-) (biofizikus) Matkó János (1952-) (biológus) Mátyus László (1956-) (biofizikus) Krasznai Zoltán (1950-) (biofizikus) Szabó Gábor (1953-) (biofizikus) Pieri, Carlo Gáspár Rezső (1944-) (biofizikus) Szöllősi János (1953-) (biofizikus)
Internet cím:DOI
elektronikus változat
Borító:

7.

001-es BibID:BIBFORM060604
035-os BibID:(scopus)0035871630 (wos)000170948300037
Első szerző:Gáspár Rezső (biofizikus)
Cím:Clustering of class I HLA oligomers with CD8 and TCR: three-dimensional models based on fluorescence resonance energy transfer and crystallographic data / Rezső Gáspár Jr., Péter Bagossi, László Bene, János Matkó, János Szöllősi, József Tőzsér, László Fésüs, Thomas A. Waldmann, Sándor Damjanovich
Dátum:2001
Megjegyzések:Fluorescence resonance energy transfer (FRET) data, in accordance with lateral mobility measurements, suggested the existence of class I HLA dimers and oligomers at the surface of live human cells, including the B lymphoblast cell line (JY) used in the present study. Intra- and intermolecular class I HLA epitope distances were measured on JY B cells by FRET using fluorophoreconjugated Ag-binding fragments of mAbs W6/32 and L368 directed against structurally well-characterized heavy and light chain epitopes, respectively. Out-of-plane location of these epitopes relative to the membrane-bound BODIPY-PC (2-(4,4-difluoro-5-(4- phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine) was also determined by FRET. Computer-simulated docking of crystallographic structures of class I HLA and epitope-specific Ag-binding fragments, with experimentally determined interepitope and epitope to cell surface distances as constraints, revealed several sterically allowed and FRET-compatible class I HLA dimeric and tetrameric arrangements. Extension of the tetrameric class I HLA model with interacting TCR and CD8 resulted in a model of a supramolecular cluster that may exist physiologically and serve as a functionally significant unit for a network of CD8-HLA-I complexes providing enhanced signaling efficiency even at low MHC-peptide concentrations at the interface of effector and APCs.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Journal of immunology. - 166 : 8 (2001), p. 5078-5086. -
További szerzők:Bagossi Péter (1966-2011) (biokémikus, vegyész) Bene László (1963-) (biofizikus) Matkó János (1952-) (biológus) Szöllősi János (1953-) (biofizikus) Tőzsér József (1959-) (molekuláris biológus, biokémikus, vegyész) Fésüs László (1947-) (orvos biokémikus) Waldmann, Thomas A. Damjanovich Sándor (1936-2017) (biofizikus)
Pályázati támogatás:T029947
OTKA
F020590
OTKA
T019372
OTKA
T030399
OTKA
T023873
OTKA
T030411
OTKA
FKFP 327/2000
Egyéb
ETT T05/102/2000
Egyéb
FKFP 0518/99
Egyéb
Internet cím:Intézményi repozitóriumban (DEA) tárolt változat
DOI
Borító:

8.

001-es BibID:BIBFORM039514
Első szerző:Matkó János (biológus)
Cím:GPI-microdomains (membrane rafts) and signaling of the multi-chain interleukin-2 receptor in human lymphoma/leukemia T cell lines / Matko, J., Bodnar, A., Vereb, G., Bene, L., Vamosi, G., Szentesi, G., Szollosi, J., Gaspar, R., Horejsi, V., Waldmann, T. A., Damjanovich, S.
Dátum:2002
ISSN:0014-2956
Megjegyzések:Subunits (alpha, beta and gamma) of the interleukin-2 receptor complex (IL-2R) are involved in both proliferative and activation-induced cell death (AICD) signaling of T cells. In addition, the signaling beta and gamma chains are shared by other cytokines (e.g. IL-7, IL-9, IL-15). However, the molecular mechanisms responsible for recruiting/sorting the alpha chains to the signaling chains at the cell surface are not clear. Here we show, in four cell lines of human adult T cell lymphoma/leukemia origin, that the three IL-2R subunits are compartmented together with HLA glycoproteins and CD48 molecules in the plasma membrane, by means of fluorescence resonance energy transfer (FRET), confocal microscopy and immuno-biochemical techniques. In addition to the beta and gamma(c) chains constitutively expressed in detergent-resistant membrane fractions (DRMs) of T cells, IL-2Ralpha (CD25) was also found in DRMs, independently of its ligand-occupation. Association of CD25 with rafts was also confirmed by its colocalization with GM-1 ganglioside. Depletion of membrane cholesterol using methyl-beta-cyclodextrin substantially reduced co-clustering of CD25 with CD48 and HLA-DR, as well as the IL-2 stimulated tyrosine-phosphorylation of STATs (signal transducer and activator of transcription). These data indicate a GPI-microdomain (raft)-assisted recruitment of CD25 to the vicinity of the signaling beta and gamma(c) chains. Rafts may promote rapid formation of a high affinity IL-2R complex, even at low levels of IL-2 stimulus, and may also form a platform for the regulation of IL-2 induced signals by GPI-proteins (e.g. CD48). Based on these data, the integrity of these GPI-microdomains seems critical in signal transduction through the IL-2R complex.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:European Journal Of Biochemistry. - 269 : 4 (2002), p. 1199-1208. -
További szerzők:Dóczy-Bodnár Andrea (1970-) (biofizikus) Vereb György (1965-) (biofizikus, orvos) Bene László (1963-) (biofizikus) Vámosi György (1967-) (biofizikus) Szentesi Gergely (1976-) (kémia-fizika tanár) Szöllősi János (1953-) (biofizikus) Gáspár Rezső (1944-) (biofizikus) Horejsi, Václav Waldmann, Thomas A. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:

9.

001-es BibID:BIBFORM023492
Első szerző:Matkó János (biológus)
Cím:Analysis of cell surface molecular distributions and cellular signaling by flow cytometry / J. Matkó, L. Mátyus, J. Szöllősi, L. Bene, A. Jenei, P. Nagy, A. Bodnár, S. Damjanovich
Dátum:1994
ISSN:1053-0509
Megjegyzések:Flow cytometry is a fast analysis and separation method for large cell populations, based on collection and processing of optical signals gained on a cell-by-cell basis. These optical signals are scattered light and fluorescence. Owing to its unique potential ofStatistical data analysis and sensitive monitoring of (micro)heterogeneities in large cell populations, flow cytometry?in combination with microscopic imaging techniques?is a powerful tool to study molecular details of cellular signal transduction processes as well. The method also has a widespread clinical application, mostly in analysis of lymphocyte subpopulations for diagnostic (or research) purposes in diseases related to the immune system. A special application of flow cytometry is the mapping of molecular interactions (proximity relationships between membrane proteins) at the cell surface, on a cell-by-cell basis. We developed two approaches to study such questions; both are based ondistance-dependent quenching of excited state fluorophores (donors) by fluorescent or dark (nitroxide radical) acceptors via Förstertype dipole-dipole resonance energy transfer (FRET) and long-range electron transfer (LRET) mechanisms, respectively. A critical evaluation of these methods using donor- or acceptor-conjugated monoclonal antibodies (or their Fab fragments) to select the appropriate cell surface receptor or antigen will be presented in comparison with other approaches for similar purposes. The applicability of FRET and LRET for two-dimensional antigen mapping as well as for detection of conformational changes in extracellular domains of membrane-bound proteins is discussed and illustrated by examples of several lymphoma cell lines. Another special application area of flow cytometry is the analysis of different aspects of cellular signal transduction, e.g., changes of intracellular ion (Ca2+, H+, Na+) concentrations, regulation of ion channel activities, or more complex physiological responses of cell to external stimuli via correlated fluorescence and scatter signal analysis, on a cell-by-cell basis. This way different signaling events such as changes in membrane permeability, membrane potential, cell size and shape, ion distribution, cell density, chromatin structure, etc., can be easily and quickly monitored over large cell populations with the advantage of revealing microheterogeneities in the cellular responses. Flow cytometry also offers the possibility to follow the kinetics of slow (minute- and hour-scale) biological processes in cell populations. These applications are illustrated by the example of complex flow cytometric analysis of signaling in extracellular ATP-triggered apoptosis (programmed cell death) of murine thymic lymphocytes.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
fluorescence
flow cytometry
energy transfer
electron transfer
protein-protein interaction
signal transduction
egyetemen (Magyarországon) készült közlemény
Megjelenés:Journal Of Fluorescence 4 : 4 (1994), p. 303-314. -
További szerzők:Mátyus László (1956-) (biofizikus) Szöllősi János (1953-) (biofizikus) Bene László (1963-) (biofizikus) Jenei Attila (1966-) (biofizikus) Nagy Péter (1971-) (biofizikus) Dóczy-Bodnár Andrea (1970-) (biofizikus) Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változa
Borító:

10.

001-es BibID:BIBFORM004878
035-os BibID:(scopus)26444586247 (wos)000232299300007
Első szerző:Nagy Péter (biofizikus)
Cím:Novel calibration method for flow cytometric fluorescence resonance energy transfer measurements between visible fluorescent proteins / Nagy, P., Bene, L., Hyun, W. C., Vereb, G., Braun, M., Antz, C., Paysan, J., Damjanovich, S., Park, J. W., Szollosi, J.
Dátum:2005
ISSN:1552-4922
Megjegyzések:The combination of fluorescence resonance energy transfer (FRET) and flow cytometry offers a statistically firm approach to study protein associations. Fusing green fluorescent protein (GFP) to a studied protein usually does not disturb the normal function of a protein, but quantitation of FRET efficiency calculated between GFP derivatives poses a problem in flow cytometry. METHODS: We generated chimeras in which cyan fluorescent protein (CFP) was separated by amino acid linkers of different sizes from yellow fluorescent protein (YFP) and used them to calibrate the cell-by-cell flow cytometric FRET measurements carried out on two different dual-laser flow cytometers. Then, CFP-Kip1 was coexpressed in yeast cells with YFP and cyclin-dependent kinase-2 (Cdk2) and served as a positive control for FRET measurements, and CFP-Kip1 coexpressed with a random peptide fused to YFP was the negative control. RESULTS: We measured donor, direct, and sensitized acceptor fluorescence intensities and developed a novel way to calculate a factor (alpha) that characterized the fluorescence intensity of acceptor molecules relative to the same number of excited donor molecules, which is essential for quantifying FRET efficiency. This was achieved by calculating FRET efficiency in two different ways and minimizing the squared difference between the two results by changing alpha. Our method reliably detected the association of Cdk2 with its inhibitor, Kip1, whereas the nonspecific FRET efficiency between Cdk2 and a random peptide was negligible. We identified and sorted subpopulations of yeast cells showing interaction between the studied proteins. CONCLUSIONS: We have described a straightforward novel calibration method to accurately quantitate FRET efficiency between GFP derivatives in flow cytometry
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
analysis
Biophysics
Calibration
Cells
chemistry
Cyclin-Dependent Kinase 2
Cyclin-Dependent Kinase Inhibitor p27
Energy Transfer
Flow Cytometry
Fluorescence
Fluorescence Resonance Energy Transfer
Hungary
metabolism
methods
Protein Binding
Proteins
Research
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Support
Megjelenés:Cytometry. Part A. - 67 : 2 (2005), p. 86-96. -
További szerzők:Bene László (1963-) (biofizikus) Hyun, William C. Vereb György (1965-) (biofizikus, orvos) Braun, Manuel Antz, Christof Paysan, Jacques Damjanovich Sándor (1936-2017) (biofizikus) Park, John W. Szöllősi János (1953-) (biofizikus)
Internet cím:elektronikus változat
DOI
Borító:

11.

001-es BibID:BIBFORM004634
035-os BibID:(scopus)0032101894 (wos)000073937700007
Első szerző:Nagy Péter (biofizikus)
Cím:EGF-induced redistribution of erbB2 on breast tumor cells : flow and image cytometric energy transfer measurements / Nagy, P., Bene, L., Balazs, M., Hyun, W. C., Lockett, S. J., Chiang, N. Y., Waldman, F., Feuerstein, B. G., Damjanovich, S., Szollosi, J.
Dátum:1998
Megjegyzések:erbB2, a member of the epidermal growth factor (EGF) receptor-type tyrosine kinase receptor family, is overexpressed in breast carcinomas with poor prognosis. We examined the cell surface association of this receptor with itself and with other cell surface proteins by the Forster-type fluorescence resonance energy transfer using whole antibodies and Fab fragments. We found that erbB2 molecules homoassociate in unstimulated SK-BR-3, BT474 and BT474-M (a metastatic version of the parent BT474 line) breast tumor cells, and that the interaction was enhanced by EGF treatment in suspensions of SK-BR-3 and BT474-M cells. BT474 cells (with low EGF receptor expression) and attached SK-BR-3 cells do not respond to EGF. Image microscopic energy transfer measurements found considerable pixel-by-pixel heterogeneity in the homoassociation state of erbB2. In accordance with the EGF-induced redistribution of erbB2, EGF receptor was found to be in close proximity to erbB2 in FRET measurements. By labeling different epitopes on erbB2 and the lipid bilayer, we were able to prepare an epitope map of erbB2 molecule. Our data suggest the existence of dynamic cell surface patterns of erbB2 and point to functions fulfilled by these molecular complexes.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
analysis
biosynthesis
Breast Neoplasms
Carcinoma
Cell Membrane
chemistry
drug effects
Energy Transfer
Epidermal Growth Factor
Female
Flow Cytometry
Fluorescence
Human
Hungary
metabolism
methods
Models
Molecular
pharmacology
Receptor
erbB-2
Receptors
Transferrin
Support
Non-U.S.Gov't
Tumor Cells,Cultured
Squamous Cell
ultrastructure
Megjelenés:Cytometry. - 32 : 2 (1998), p. 120-131. -
További szerzők:Bene László (1963-) (biofizikus) Balázs Margit (1952-) (sejtbiológus, molekuláris genetikus) Hyun, William C. Lockett, Steven J. Chiang, Nancy Y. Waldman, Frederick Feuerstein, Burt G. Damjanovich Sándor (1936-2017) (biofizikus) Szöllősi János (1953-) (biofizikus)
Internet cím:elektronikus változat
DOI
Borító:

12.

001-es BibID:BIBFORM046304
Első szerző:Szöllősi János (biofizikus)
Cím:Supramolecular complexes of MHC class I, MHC class II, CD20, and tetraspan molecules (CD53, CD81, and CD82) at the surface of a B cell line JY / Szollosi J., Horejsi V., Bene L., Angelisova P., Damjanovich S.
Dátum:1996
Megjegyzések:The results of previous biochemical studies indicated that a fraction of MHC class II proteins is associated with four proteins of the tetraspan family, CD37, CD53, CD81, and CD82, and possibly with other membrane components, at the surface of JY B lymphoma cells. In the present communication we used a biophysical technique, namely the flow cytometric energy transfer method, to demonstrate the proximity of these molecules at the surface of the cells. Significant energy transfer (and, therefore, proximity within the 2-10 nm range) was observed between fluorescently labeled mAbs to DR, DQ, and the tetraspan molecules CD53, CD81, and CD82. Moreover, two other B cell surface molecules, CD20 and MHC class I, were found to be close to each other and to MHC class II and the tetraspan proteins, based on the observed high energy transfer efficiencies between the relevant fluorescently labeled mAbs. The character of simultaneous energy transfer from CD20, CD53, CD81, and CD82 to DR suggests that all these molecules are in a single complex with the DR molecules (or a complex of several DR molecules) rather than that each of them is separately associated with different DR molecules. Based on these data and previous biochemical results, a model is proposed predicting that the B cell membrane contains multicomponent supramolecular complexes consisting of at least two MHC class I and at least one DR, DQ, CD20, CD53, CD81, and CD82 molecules. Closer analysis of the energy transfer efficiencies makes it possible to suggest mutual orientations of the components within the complex. Participation of other molecules, not examined in this study (CD19 and CD37), in these supramolecular structures cannot be ruled out. These large assemblies of multiple B cell surface molecules may play a role in signaling through MHC molecules and in Ag presentation to T cells.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:The Journal of Immunology. - 157 : 7 (1996), p. 2939-2946. -
További szerzők:Horejsi, Václav Bene László (1963-) (biofizikus) Angelisova, P. Damjanovich Sándor (1936-2017) (biofizikus)
Internet cím:Szerző által megadott URL
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