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001-es BibID:	BIBFORM069692
Első szerző:	Ungvári Tamás
Cím:	Perrin and Förster unified : dual-laser triple-polarization FRET (3polFRET) for interactions at the Förster-distance and beyond / Tamás Ungvári, Péter Gogolák, Miklós Bagdány, László Damjanovich, László Bene
Dátum:	2016
ISSN:	0167-4889
Megjegyzések:	Dual laser flow cytometric energy transfer (FCET)--elaborated by Trón et al. in 1984--is an efficient and rapid way of measuring FRET on large cell populations. FRET efficiency and the donor and acceptor concentrations are determined from one donor and two acceptor signals. In this communication this method is extended towards the domain of receptor dynamics by the detection of polarized components of the three intensities. By enabling a complete description of the proximity and dynamics of FRET-systems, the new measuring scheme allows a more refined description of both the structure and dynamics of cell surface receptor clusters at the nano-scale and beyond. Associated donor fraction, limiting anisotropy and rotational correlation time of the donor, acceptor anisotropy and cell-by-cell estimation of the orientation factor for FRET (K2) are available in the steady state on a single FRET sample in a very rapid and statistically efficient way offered by flow cytometry. For a more sensitive detection of conformational changes the "polarized FRET indices"--quantities composed from FRET efficiency and anisotropies--are proposed. The method is illustrated by measurements on a FRET system with changing FRET-fraction and on a two donor-one acceptor-system, when the existence of receptor trimers are proven by the detection of "hetero-FRET induced homo-FRET relief", i.e. the diminishing of homo-FRET between the two donors in the presence of a donor quencher. The method also offers higher sensitivity for assessing conformational changes at the nano-scale, due to its capability for the simultaneous detection of changes of proximity and relative orientations of the FRET donor and acceptor. Although the method has been introduced in the context of FRET, it is more general: It can be used for monitoring triple-anisotropy correlations also in those cases when FRET actually does not occur, e.g. for interactions occuring beyond the Förster-distance R0. Interpretation of K2 has been extended.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Triple-anisotropy correlations Donor anisotropy Acceptor anisotropy Orientation factor for FRET Homo-FRET relief FRET-fraction
Megjelenés:	Biochimica et Biophysica Acta (BBA). Molecular Cell Research. - 1863 : 4 (2016), p. 703-716. -
További szerzők:	Gogolák Péter (1968-) (biológus, immunológus) Bagdány Miklós Damjanovich László (1960-) (általános sebész) Bene László (1963-) (biofizikus)
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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