CCL

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1.

001-es BibID:BIBFORM066281
Első szerző:Balogh Enikő (molekuláris biológus)
Cím:Impaired Immunosuppressive Effect of Bronchoalveolar Mesenchymal Stem Cells in Hypersensitivity Pneumonitis : preliminary findings / Eniko Balogh, Bela Nagy Jr., Agnes Gyetvai, Zsolt Bene, Zoltan Hendrik, Viktoria Jeney, Peter Nagy, Agnes Papp, Jozsef Balla, Gyorgy Balla, Janos Kappelmayer, Bela Nagy
Dátum:2018
ISSN:1552-4949
Megjegyzések:Bronchoalveolar mesenchymal stem cells (MSCs) play an important role in the maintenance of lung integrity. Therapeutic application of bone marrow-derived MSCs reduced chronic bronchial inflammation in idiopathic pulmonary fibrosis, and improved the ratio of survivors in sepsis with pneumonia. This study investigated the effect of MSCs from bronchoalveolar lavage fluid (BALF) of hypersensitivity pneumonitis (HP) on T-cell function under in vitro conditions.METHODS:Bronchoalveolar MSCs were obtained via bronchoscopy with BAL from children with severe subacute HP. As control, BALF MSCs were assessed from children without any inflammatory lung disease. Isolated MSCs were characterized via immunophenotyping by flow cytometry and confocal laser scanning microscopy. HP-derived and healthy separated peripheral blood mononuclear cells (PBMCs) were stimulated by 5 ?g/mL phytohemagglutinin in the presence of HP-derived or control MSCs in 5-day cultures. Proliferation and activation of T-cells were characterized by the mean fluorescence intensity (MFI) of 5,6-carboxyfluorescein-diacetat succinimidyl ester (CFSE) and CD25, CD69 as well as HLA-DR surface positivities, respectively.RESULTS:HP-derived MSCs showed significantly lower level of CD73, CD90, and CD105 expression compared to control MSCs in both flow cytometric and confocal microscopic experiments. MSCs from HP did not reduce T-cell proliferation based on CFSE MFI values, while the level of CD25 expression on both control and HP-derived CD4+ and CD8+ T-cells was significantly reduced by normal MSCs, while HP-derived MSCs did not have any significant effect. The level of other activation markers was not markedly modulated by MSCs.CONCLUSIONS:BALF MSCs from HP are unable to downregulate the proliferation and activation of T-cells that may support the development of recurrent intrapulmonary inflammation in HP. ? 2016 Clinical Cytometry Society.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
T-lymphocyte proliferation and activation
bronchoalveolar lavage
hypersensitivity pneumonitis
mesenchymal stem cells
Megjelenés:Cytometry Part B-Clinical Cytometry. - 94 : 2 (2018), p. 363-368. -
További szerzők:Nagy Béla Jr. (1980-) (labordiagnosztikai szakorvos) Gyetvai Ágnes Bene Zsolt (1981-) (orvos) Hendrik Zoltán (1986-) (orvos) Jeney Viktória (1971-) (vegyész, kémia tanár) Nagy Péter (1971-) (biofizikus) Papp Ágnes (1967-) (gyermekgyógyász, pulmonológus) Balla József (1959-) (belgyógyász, nephrológus) Balla György (1953-) (csecsemő és gyermekgyógyász, neonatológus) Kappelmayer János (1960-) (laboratóriumi szakorvos) Nagy Béla (1949-) (csecsemő- és gyermekgyógyász, gyermek-tüdőgyógyász)
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2.

001-es BibID:BIBFORM113506
035-os BibID:(scopus)85150638841 (wos)000948192400001
Első szerző:Batta Ágnes
Cím:Improved estimation of the ratio of detection efficiencies of excited acceptors and donors for measurements / Batta Ágnes, Hajdu Tímea, Nagy Peter
Dátum:2023
ISSN:1552-4922 1552-4930
Megjegyzések:Forster resonance energy transfer (FRET) is a radiationless interaction between a donor and an acceptor whose distance dependence makes it a sensitive tool for studying the oligomerization and the structure of proteins. When FRET is determined by measuring the sensitized emission of the acceptor, a parameter characterizing the ratio of detection efficiencies of an excited acceptor versus an excited donor is invariably involved in the formalism. For FRET measurements involving fluorescent antibodies or other external labels, this parameter, designated by alpha, is usually determined by comparing the intensity of a known number of donors and acceptors in two independent samples leading to a large statistical variability if the sample size is small. Here, we present a method that improves precision by applying microbeads with a calibrated number of antibody binding sites and a donor-acceptor mixture in which donors and acceptors are present in a certain, experimentally determined ratio. A formalism is developed for determining alpha and the superior reproducibility of the proposed method compared to the conventional approach is demonstrated. Since the novel methodology does not require sophisticated calibration samples or special instrumentation, it can be widely applied for the quantification of FRET experiments in biological research.
Tárgyszavak:Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry Part A. - 103 : 7 (2023), p. 563-574. -
További szerzők:Hajdu Tímea (1991-) (biomérnök) Nagy Péter (1971-) (biofizikus)
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3.

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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Intézményi repozitóriumban (DEA) tárolt változat
Borító:

4.

001-es BibID:BIBFORM004864
035-os BibID:(scopus)26444452209 (wos)000232299300016
Első szerző:Friedländer Elza (biofizikus)
Cím:Signal transduction of erbB receptors in trastuzumab (Herceptin) sensitive and resistant cell lines : local stimulation using magnetic microspheres as assessed by quantitative digital microscopy / Friedlander, E., Arndt-Jovin, D. J., Nagy, P., Jovin, T. M., Szollosi, J., Vereb, G.
Dátum:2005
ISSN:1552-4922
Megjegyzések:ErbB2 (HER-2), a member of the epidermal growth factor (EGF) receptor family, is a class I transmembrane receptor tyrosine kinase. Although erbB2 has no known physiologic ligand, it can form complexes with other members of the family and undergo transactivation of its very potent kinase activity, thereby initiating downstream signaling and cell proliferation. ErbB2 is a frequent pathologic marker in ductal invasive breast carcinomas and is targeted by using a specific humanized monoclonal antibody, trastuzumab (Herceptin). The antibody is effective in only 20% to 50% of erbB2-positive tumors, and this resistance, as yet poorly understood, constitutes a major therapeutic challenge. METHODS: Magnetic microspheres coated with ligands or antibodies are widely used for separation of proteins and cells and allow localized, high intensity, and precisely timed stimulation of cells. We used EGF- and trastuzumab-covered paramagnetic microspheres, quantitative confocal laser scanning microscopy, and digital image processing to investigate the (trans)activation of and local signal propagation from erbB1 and erbB2 on trastuzumab sensitive and resistant carcinoma cell lines expressing these receptors at high levels. RESULTS: On A431 cells expressing high levels of endogenous erbB1 and transfected erbB2-mYFP (A4-erbB2-mYFP F4 cell line), EGF-coupled-microspheres activated erbB1 and transactivated erbB2-mYFP. In two other cell lines with comparable erbB2 expression but lower levels of erbB1, EGF microspheres transactivated erbB2 less efficiently. Trastuzumab in solution activated erbB2 on A4-erbB2-mYFP and the trastuzumab sensitive SKBR-3 cells, but only negligibly on the resistant JIMT-1 cells that showed a 10 times higher K(d) for the antibody. Nevertheless, pronounced erbB2 activation and tyrosine phosphorylation could be detected after stimulation with trastuzumab-coupled microspheres in all cell lines, although transactivation of erbB1 was negligible. Receptor phosphorylation was restricted to the immediate proximity of the microspheres, i.e., receptor clusters external to these locations remained inactive. CONCLUSION: ErbB1 ligand and erbB2 specific antibody attached to magnetic microspheres are efficient tools in assessing erbB activation, localized signal propagation, and erbB heterodimer formation. Trastuzumab coupled to microspheres is more efficient at accessing erbB2 and activating it than trastuzumab in solution
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Animals
Antibodies
Antibodies,Monoclonal
Biophysics
Carcinoma
Cell Line
Cell Line,Tumor
Cell Proliferation
Cells
chemistry
drug effects
Drug Resistance,Neoplasm
Epidermal Growth Factor
genetics
Humans
Hungary
Ligands
Magnetics
metabolism
methods
Microscopy
Microspheres
pharmacology
Phosphorylation
Proteins
Receptor,erbB-2
Research
Signal Transduction
Solubility
Support
Trans-Activation (Genetics)
Megjelenés:Cytometry. Part A. - 67 : 2 (2005), p. 161-171. -
További szerzők:Arndt-Jovin, Donna J. Nagy Péter (1971-) (biofizikus) Jovin, Thomas M. Szöllősi János (1953-) (biofizikus) Vereb György (1965-) (biofizikus, orvos)
Internet cím:elektronikus változat
DOI
Borító:

5.

001-es BibID:BIBFORM005185
035-os BibID:(scopus)46449085498 (wos)000257176000003
Első szerző:Kiss Endre (Budapest)
Cím:Cytometry of raft and caveola membrane microdomains : from flow and imaging techniques to high throughput screening assays / Kiss, E., Nagy, P., Balogh, A., Szollosi, J., Matko, J.
Dátum:2008
ISSN:552-4930 (Electronic)
Megjegyzések:The evolutionarily developed microdomain structure of biological membranes has gained more and more attention in the past decade. The caveolin-free "membrane rafts," the caveolin-expressing rafts (caveolae), as well as other membrane microdomains seem to play an essential role in controlling and coordinating cell-surface molecular recognition, internalization/endocytosis of the bound molecules or pathogenic organisms and in regulation of transmembrane signal transduction processes. Therefore, in many research fields (e.g. neurobiology and immunology), there is an ongoing need to understand the nature of these microdomains and to quantitatively characterize their lipid and protein composition under various physiological and pathological conditions. Flow and image cytometry offer many sophisticated and routine tools to study these questions. In this review, we give an overview of the past efforts to detect and characterize these membrane microdomains by the use of classical cytometric technologies, and finally we will discuss the results and perspectives of a new line of raft cytometry, the "high throughput screening assays of membrane microdomains," based on "lipidomic" and "proteomic" approaches.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Animals
article
Caveolae
Cell Membrane
chemistry
Detergents
Flow Cytometry
Fluorescence Resonance Energy Transfer
Humans
Hungary
Image Cytometry
immunology
Lipids
Membrane Lipids
Membrane Microdomains
metabolism
methods
pharmacology
Protein Structure, Tertiary
Proteomics
Research
Research Support
Signal Transduction
Support
Megjelenés:Cytometry. Part A. - 73A : 7 (2008), p. 599-614. -
További szerzők:Nagy Péter (1971-) (biofizikus) Balogh Andrea Szöllősi János (1953-) (biofizikus) Matkó János (1952-) (biológus)
Internet cím:DOI
elektronikus változat
Borító:

6.

001-es BibID:BIBFORM010354
035-os BibID:(scopus)67651042934 (wos)000268289300002
Első szerző:Kovács Tamás (általános orvos)
Cím:The Density of GM1-Enriched Lipid Rafts Correlates Inversely with the Efficiency of Transfection Mediated by Cationic Liposomes / Kovacs, T., Karasz, A., Szollosi, J., Nagy, P.
Dátum:2009
ISSN:1552-4922 (Print)
Megjegyzések:Although cationic liposome-mediated transfection has become a standard procedure, the mechanistic details of the process are unknown. It has been suggested that endocytic uptake of lipoplexes is efficient, and transfectability is largely determined by later steps. In this article, we stained GM1-enriched membrane microdomains, a subclass of lipid rafts, with subunit B of cholera toxin and correlated transfection efficiency with their density by quantitatively evaluating microscopic images. We found a strong anticorrelation between the density of GM1-enriched membrane microdomains and the efficacy of transfection monitored by measuring the expression level of GFP in different cell lines transfected by lipofection using two different transfection agents. These findings imply that GM1-enriched membrane microdomains interfere with the process of lipofection. The blocked step must be endocytosis since the accumulation of fluorescently labeled plasmids was lower in cells with high content of GM1-enriched membrane microdomains. Such a correlation was not observed in cells transfected by electroporation. By comparing the efficiency of lipofection in several cell lilies we found that those with a high density of GM1-enriched membrane microdomains were the most resistant to transfection. We conclude that the inhibition of lipofection by GM1-enriched membrane microdomains is a general rule, and that endocytosis of lipoplexes call be rate limiting in cells with high density of GM1-enriched membrane rafts.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
article
Cell Line
Cells
Cholera Toxin
Endocytosis
Membrane Microdomains
Plasmids
Transfection
Megjelenés:Cytometry. Part A. - 75A : 8 (2009), p. 650-657. -
További szerzők:Kárász Andrea (biofizikus) Szöllősi János (1953-) (biofizikus) Nagy Péter (1971-) (biofizikus)
Internet cím:DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:

7.

001-es BibID:BIBFORM070072
035-os BibID:(WoS)000405907100005 (Scopus)85023170074
Első szerző:Mocanu, Maria-Magdalena
Cím:Detection of protein interactions by Subcellular Localization Assay / Mocanu Maria-Magdalena, Nagy Péter, Szöllősi János
Dátum:2017
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry Part A. - 91 : 7 (2017), p. 657-658. -
További szerzők:Nagy Péter (1971-) (biofizikus) Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:GINOP-2.3.2-15-2016-00050
GINOP
GINOP-2.3.2-15-2016-00020
GINOP
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Intézményi repozitóriumban (DEA) tárolt változat
Borító:

8.

001-es BibID:BIBFORM071351
035-os BibID:(scopus)84963800113 (wos)000374730400007
Első szerző:Nagy Péter (biofizikus)
Cím:rFRET : a comprehensive, Matlab-based program for analyzing intensity-based ratiometric microscopic FRET experiments / Nagy Peter, Szabó Ágnes, Váradi Tímea, Kovács Tamás, Batta Gyula, Szöllősi János
Dátum:2016
ISSN:1552-4922 1552-4930
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry Part A. - 89 : 4 (2016), p. 376-384. -
További szerzők:Nagyné Szabó Ágnes Timea (1982-) (vegyész) Váradi Tímea (1982-) (okleveles vegyész) Kovács Tamás (1985-) (általános orvos) Batta Gyula (1979-) (biológus) Szöllősi János (1953-) (biofizikus)
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Intézményi repozitóriumban (DEA) tárolt változat
Borító:

9.

001-es BibID:BIBFORM057540
035-os BibID:(scopus)84911424770 (wos)000344166300007
Első szerző:Nagy Péter (biofizikus)
Cím:Maximum likelihood estimation of FRET efficiency and its implications for distortions in pixelwise calculation of FRET in microscopy / Peter Nagy, Ágnes Szabó, Tímea Váradi, Tamás Kovács, Gyula Batta, János Szöllősi
Dátum:2014
ISSN:1552-4922 1552-4930
Megjegyzések:Ratiometric determination of the efficiency of fluorescence or Forster resonance energy € transfer (FRET) is one of the most widespread methods for the characterization of protein clustering and conformation. Low photon numbers, often present in pixel-by-pixel determination of FRET efficiency in digital microscopy, result in large uncertainties in the derived FRET parameter. Here, we propose a method based on maximum likelihood estimation (MLE) of FRET efficiency using photon counting detectors to overcome this limitation. Intensities measured in the donor, FRET, and acceptor channels were all assumed to follow Poisson statistics as a result of detector shot noise. The joint probability of photon numbers detected in the donor, FRET, and acceptor channels was derived using an equation describing the relationship between the three measured intensities. The FRET efficiency generating the measured photon numbers with the largest likelihood was determined iteratively providing a single FRET value for all pixels in the calculation. Since as few as 100 pixels are sufficient to provide a maximum likelihood estimate for FRET, biological variability in FRET values can be revealed by performing the analysis for regions of interests in an image. Since the algorithm provides the probability of a combination of donor, FRET, and acceptor intensities observed in each individual pixel given a certain FRET efficiency, outlier pixels with low probabilities could be excluded from the analysis. Simulations carried out with low photon numbers in the presence and absence of outlier pixels revealed that the proposed approach can reliably and reproducibly estimate FRET efficiency. In addition, systematic evaluation of the simulation results showed that the distribution of pixel-by-pixel FRET efficiencies is skewed, and the mean of these FRET values is a biased and unreliable estimate of the FRET efficiency. In the absence of outlier pixels, FRET calculated from summed donor, FRET, and acceptor intensities proved to be as reliable as MLE. We conclude that MLE of FRET outperforms calculations using summed and pixel-bypixel intensities in biologically relevant situations involving low photon numbers and outlier pixels.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
FRET
maximum likelihood estimation (MLE)
microscopy
Poisson statistics
error propagation
Megjelenés:Cytometry Part A. - 85 : 11 (2014), p. 942-952. -
További szerzők:Nagyné Szabó Ágnes Timea (1982-) (vegyész) Váradi Tímea (1982-) (okleveles vegyész) Kovács Tamás (1985-) (általános orvos) Batta Gyula (1979-) (biológus) Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:TAMOP-4.2.2.A-11/1/KONV-2012-0025
TÁMOP
OTKA-K103906
OTKA
OTKA-NK101337
OTKA
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Intézményi repozitóriumban (DEA) tárolt változat
Borító:

10.

001-es BibID:BIBFORM030392
035-os BibID:(scopus)84856214641 (wos)000299376900004
Első szerző:Nagy Péter (biofizikus)
Cím:How to avoid bleeding in Forster resonance energy transfer / Nagy, P., Szollosi, J.
Dátum:2012
Tárgyszavak:Orvostudományok Elméleti orvostudományok hozzászólás
folyóiratcikk
Biophysics
Energy Transfer
ENERGY-TRANSFER
Hungary
resonance energy transfer
Megjelenés:Cytometry. Part A. - 81A : 2 (2012), p. 108-109. -
További szerzők:Szöllősi János (1953-) (biofizikus)
Internet cím:DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:

11.

001-es BibID:BIBFORM010357
035-os BibID:(scopus)70349515373 (wos)000270419700002
Első szerző:Nagy Péter (biofizikus)
Cím:Proximity or no proximity : that is the question-but the answer is more complex / Nagy, P., Szollosi, J.
Dátum:2009
ISSN:1552-4922 (Print)
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Cytometry. Part A. - 75A : 10 (2009), p. 813-815. -
További szerzők:Szöllősi János (1953-) (biofizikus)
Internet cím:DOI
Intézményi repozitóriumban (DEA) tárolt változat
Borító:

12.

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)
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