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001-es BibID:	BIBFORM119122
035-os BibID:	(WoS)001176727000001 (Scopus)85186901297
Első szerző:	Pethő Zoltán (orvos)
Cím:	A synthetic flavonoid derivate in the plasma membrane transforms the voltage-clamp fluorometry signal of CiHv1 / Zoltán Pethő, Dávid Pajtás, Martina Piga, Zsuzsanna Magyar, Florina Zakany, Tamas Kovacs, Nace Zidar, Gyorgy Panyi, Zoltan Varga, Ferenc Papp
Dátum:	2024
ISSN:	1742-464X
Megjegyzések:	Voltage-clamp fluorometry (VCF) enables the study of voltage-sensitive proteins through fluorescent labeling accompanied by ionic current measurements for voltage-gated ion channels. The heterogeneity of the fluorescent signal represents a significant challenge in VCF. The VCF signal depends on where the cysteine mutation is incorporated, making it difficult to compare data among different mutations and different studies and standardize their interpretation. We have recently shown that the VCF signal originates from quenching amino acids in the vicinity of the attached fluorophores, together with the effect of the lipid microenvironment. Based on these, we performed experiments to test the hypothesis that the VCF signal could be altered by amphiphilic quenching molecules in the cell membrane. Here we show that a phenylalanine-conjugated flavonoid (4-oxo-2-phenyl- 4H-chromene-7-yl)-phenylalanine, (later Oxophench) has potent effects on the VCF signals of the Ciona intestinalis HV1 (CiHv1) proton channel. Using spectrofluorimetry, we showed that Oxophench quenches TAMRA (5(6)-carboxytetramethylrhodamine-(methane thiosulfonate)) fluorescence. Moreover, Oxophench reduces the baseline fluorescence in oocytes and incorporates into the cell membrane while reducing the membrane fluidity of HEK293 cells. Our model calculations confirmed that Oxophench, a potent membrane-bound quencher, modifies the VCF signal during conformational changes. These results support our previously published model of VCF signal generation and point out that a change in the VCF signal may not necessarily indicate an altered conformational transition of the investigated protein.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk CiHv1 flavonoid derivate fluorescencequenching membrane fluidity voltageclamp fluorometry
Megjelenés:	Febs Journal. - [Epub ahead of print] (2024). -
További szerzők:	Pajtás Dávid (1987-) (vegyész) Piga Martina Magyar Zsuzsanna (1981-) (orvos) Zákány Florina (1989-) (általános orvos) Kovács Tamás (1985-) (általános orvos) Zidar, Nace Panyi György (1966-) (biofizikus) Varga Zoltán (1969-) (biofizikus, szakfordító) Papp Ferenc (1979-) (biofizikus)
Pályázati támogatás:	132906 OTKA 2019-2.1.11-TÉT-2019-00059 Egyéb BO/00355/21/8 Bolyai MTA ÚNKP-22-5-DE-420 Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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