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001-es BibID:	BIBFORM065088
035-os BibID:	(Cikkazonosító)33022 (WOS)000382917700001 (Scopus)85047916061
Első szerző:	Vámosi György (biofizikus)
Cím:	EGFP oligomers as natural fluorescence and hydrodynamic standards / György Vámosi, Norbert Mücke, Gabriele Müller, Jan Wolfgang Krieger, Ute Curth, Jörg Langowski, Katalin Tóth
Dátum:	2016
ISSN:	2045-2322
Megjegyzések:	EGFP oligomers are convenient standards for experiments on fluorescent protein-taggedbiomolecules. In this study, we characterized their hydrodynamic and fluorescence properties.Diffusion coefficients D of EGFP1-4 were determined by analytical ultracentrifugation withfluorescence detection and by fluorescence correlation spectroscopy (FCS), yielding83.4?48.2 ?m2/s and 97.3?54.8 ?m2/s from monomer to tetramer. A "barrels standing in arow" model agreed best with the sedimentation data. Oligomerization red-shifted EGFPemission spectra without any shift in absorption. Fluorescence anisotropy increased,indicating homoFRET between the subunits. Fluorescence lifetime decreased only slightly(4%) indicating insignificant quenching by FRET to subunits in non-emitting states. FCSmeasuredD, particle number and molecular brightness depended on dark states and lightinducedprocesses in distinct subunits, resulting in a dependence on illumination powerdifferent for monomers and oligomers. Since subunits may be in "on" (bright) or "off" (dark)states, FCS-determined apparent brightness is not proportional to that of the monomer. Fromits dependence on the number of subunits, the probability of the "on" state for a subunit wasdetermined to 96% at pH8 and 77% at pH6.38, i.e., protonation increases the dark state. Thesefluorescence properties of EGFP oligomeric standards can assist interpreting results fromoligomerized EGFP fusion proteins of biological interest.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk oligo-EGFP FP photophysics diffusion coefficient molecular brightness protein oligomerization
Megjelenés:	Scientific Reports. - 6 (2016), p. 1-12. -
További szerzők:	Mücke, Norbert Müller, Gabriele Krieger, Jan Wolfgang Curth, Ute Langowski, Jörg Tóth Katalin (Heidelberg)
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat
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