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001-es BibID:	BIBFORM079931
035-os BibID:	(cikkazonosító)3370 (scopus)85071327332 (wos)000477041100254
Első szerző:	Vámosi György (biofizikus)
Cím:	EGF Receptor Stalls upon Activation as Evidenced by Complementary Fluorescence Correlation Spectroscopy and Fluorescence Recovery after Photobleaching Measurements / Vámosi György, Friedländer-Brock Elza, Ibrahim Shehu M., Brock Roland, Szöllősi János, Vereb György
Dátum:	2019
ISSN:	1661-6596 1422-0067
Megjegyzések:	To elucidate the molecular details of the activation-associated clustering of epidermal growth factor receptors (EGFRs), the time course of the mobility and aggregation states of eGFP tagged EGFR in the membranes of Chinese hamster ovary (CHO) cells was assessed by in situ mobility assays. Fluorescence correlation spectroscopy (FCS) was used to probe molecular movements of small ensembles of molecules over short distances and time scales, and to report on the state of aggregation. The diffusion of larger ensembles of molecules over longer distances (and time scales) was investigated by fluorescence recovery after photobleaching (FRAP). Autocorrelation functions could be best fitted by a two-component diffusion model corrected for triplet formation and blinking. The slow, 100?1000 ms component was attributed to membrane localized receptors moving with free Brownian diffusion, whereas the fast, ms component was assigned to cytosolic receptors or their fragments. Upon stimulation with 50 nM EGF, a significant decrease from 0.11 to 0.07 ?m2/s in the diffusion coefficient of membrane-localized receptors was observed, followed by recovery to the original value in ~20 min. In contrast, the apparent brightness of diffusing species remained the same. Stripe FRAP experiments yielded a decrease in long-range molecular mobility directly after stimulation, evidenced by an increase in the recovery time of the slow component from 13 to 21.9 s. Our observations are best explained by the transient attachment of ligand-bound EGFRs to immobile or slowly moving structures such as the cytoskeleton or large, previously photobleached receptor aggregates.
Tárgyszavak:	Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban Fluorescence correlation spectroscopy FCS fluorescence recovery after photobleaching FRAP epidermal growth factor receptor translational diffusion EGFR?eGFP fusion protein
Megjelenés:	International Journal Of Molecular Sciences. - 20 : 13 (2019), p. 1-22. -
További szerzők:	Friedländer Elza (1980-) (biofizikus) Ibrahim, Shehu M. Brock Roland Szöllősi János (1953-) (biofizikus) Vereb György (1965-) (biofizikus, orvos)
Pályázati támogatás:	K119690 OTKA NN129371 OTKA GINOP-2.3.2-15-2016-00050 GINOP GINOP-2.3.3-15-2016-00003 GINOP
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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