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001-es BibID:BIBFORM062699
035-os BibID:(scopus)84925853989 (wos)000354040400007
Első szerző:Shrestha, Dilip (biológus)
Cím:Understanding FRET as a Research Tool for Cellular Studies / Dilip Shrestha, Attila Jenei, Péter Nagy, György Vereb, János Szöllősi
Dátum:2015
ISSN:1661-6596 1422-0067
Megjegyzések:Communication of molecular species through dynamic association and/or dissociation at various cellular sites governs biological functions. Understanding these physiological processes require delineation of molecular events occurring at the level of individual complexes in a living cell. Among the few non-invasive approaches with nanometer resolution are methods based on Förster Resonance Energy Transfer (FRET). FRET is effective at a distance of 1?10 nm which is equivalent to the size of macromolecules, thus providing an unprecedented level of detail on molecular interactions. The emergence of fluorescent proteins and SNAP- and CLIP- tag proteins provided FRET with the capability to monitor changes in a molecular complex in real-time making it possible to establish the functional significance of the studied molecules in a native environment. Now, FRET is widely used in biological sciences, including the field of proteomics, signal transduction, diagnostics and drug development to address questions almost unimaginable with biochemical methods and conventional microscopies. However, the underlying physics of FRET often scares biologists. Therefore, in this review, our goal is to introduce FRET to non-physicists in a lucid manner. We will also discuss our contributions to various FRET methodologies based on microscopy and flow cytometry, while describing its application for determining the molecular heterogeneity of the plasma membrane in various cell types.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
FRET
Megjelenés:International Journal Of Molecular Sciences. - 16 : 4 (2015), p. 6718-6756. -
További szerzők:Jenei Attila (1966-) (biofizikus) Nagy Péter (1971-) (biofizikus) Vereb György (1965-) (biofizikus, orvos) Szöllősi János (1953-) (biofizikus)
Pályázati támogatás:MTA-DE
MTA
Sejtbiológiai és Jelátvitel Kutatócsoport
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2.

001-es BibID:BIBFORM048515
Első szerző:Shrestha, Dilip (biológus)
Cím:CD1d favors MHC neighborhood, GM1 ganglioside proximity and low detergent sensitive membrane regions on the surface of B lymphocytes / Dilip Shrestha, Mark A. Exley, György Vereb, János Szöllősi, Attila Jenei
Dátum:2014
ISSN:0304-4165
Megjegyzések:BACKGROUND: Cluster of differentiation 1 (CD1) represents a family of proteins which is involved in lipid-based antigen presentation. Primarily, antigen presenting cells, like B cells, express CD1 proteins. Here, we examined the cell-surface distribution of CD1d, a subtype of CD1 receptors, on B lymphocytes. METHODS: Fluorescence labeling methods, including fluorescence resonance energy transfer (FRET), were employed to investigate plasma membrane features of CD1d receptors. RESULTS: High FRET efficiency was observed between CD1d and MHC I heavy chain (MHC I-HC), beta2-microglobulin (beta2m) and MHC II proteins in the plasma membrane. In addition, overexpression of CD1d reduced the expression of MHC II and increased the expression of MHC I-HC and beta2m proteins on the cell-surface. Surprisingly, beta2m dependent CD1d isoform constituted only ~15% of the total membrane CD1d proteins. Treatment of B cells with methyl-beta-cyclodextrin (MbetaCD) / simvastatin caused protein rearrangement; however, FRET demonstrated only minimal effect of these chemicals on the association between CD1d and GM1 ganglioside on cell-surface. Likewise, a modest effect was only observed in a co-culture assay between MbetaCD/simvastatin treated C1R-CD1d cells and invariant natural killer T cells on measuring secreted cytokines (IFNgamma and IL4). Furthermore, CD1d rich regions were highly sensitive to low concentration of Triton X-100. Physical proximity between CD1d, MHC and GM1 molecules was also detected in the plasma membrane. CONCLUSIONS: An intricate relationship between CD1d, MHC, and lipid species were found on the membrane of human B cells. General significance Organization of CD1d on the plasma membrane might be critical for its biological functions.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
CD1d
MHC
FRET
Rafts
Simvastatin
Megjelenés:Biochimica et Biophysica Acta (BBA). General Subjects. - 1840 : 1 (2014), p. 667-680. -
További szerzők:Exley, Mark A. (1961-) (immunológus) Vereb György (1965-) (biofizikus, orvos) Szöllősi János (1953-) (biofizikus) Jenei Attila (1966-) (biofizikus)
Internet cím:Szerző által megadott URL
DOI
Intézményi repozitóriumban (DEA) tárolt változat
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