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001-es BibID:	BIBFORM076187
035-os BibID:	(WoS)000453309600001 (Scopus)85058790521
Első szerző:	Garda Tamás (biológus)
Cím:	Allyl-isothiocyanate and microcystin-LR reveal the protein phosphatase mediated regulation of metaphase-anaphase transition in Vicia faba / Tamás Garda, Zoltán Kónya, Csongor Freytag, Ferenc Erdődi, Sándor Gonda, Gábor Vasas, Boglárka Szücs, Márta M-Hamvas, Attila Kiss-Szikszai, György Vámosi, Csaba Máthé
Dátum:	2018
ISSN:	1664-462X
Megjegyzések:	Horseradish allyl isothiocyanate (AITC, a volatile oil) and cyanobacterial microcystin-LR (MCY-LR, a cyclic heptapeptide) affect eukaryotic cell cycle. MCY-LR inhibits protein phosphatases PP1 and PP2A. We aimed to reveal the mechanisms of their cellular effects in a model eukaryote, Vicia faba. We have shown for the first time that AITC had minor effects on PP1 and PP2A activities in vitro, but it inhibited significantly PP1 in vivo. The combination of 10 ?M AITC with 10 ?M MCY-LR induced metaphase arrest after short-term (12 h) treatments. 10 ?M AITC, 0.2-10 ?M MCY-LR and their combinations induced histone H3 hyperphosphorylation, associated with the regulation of metaphase-anaphase transition. This hyperphosphorylation event occurred at any treatment which led to the inhibition of PP1 activity. 10 ?M AITC + 10 ?M MCY-LR increased the frequency of metaphase spindle anomalies, associated with metaphase arrest. We provide new insights into the mechanisms of metaphase-anaphase transition. Metaphase arrest is induced at the concomitant hyperphosphorylation of histone H3, alteration of metaphase spindle assembly and strong inhibition of PP1+PP2A activity. Near-complete blocking of metaphase-anaphase transition by rapid protein phosphatase inhibition is shown here for the first time in plants, confirming a crucial role of serine-threonine phosphatases in this checkpoint of cell cycle regulation. Tissue-dependent differences in PP1 and PP2A activities induced by AITC and MCY-LR suggest that mainly regulatory subunits are affected. AITC is a potential tool for the study of protein phosphatase function and regulation. We raise the possibility that one of the biochemical events occurring during AITC release upon wounding is the modulation of protein phosphatase dependent signal transduction pathways during the plant defence response.
Tárgyszavak:	Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Frontiers in Plant Science. - 9 (2018), p. 1-15. -
További szerzők:	Kónya Zoltán (1986-) (molekuláris biológus, biokémikus) Freytag Csongor (1993-) (biológus) Erdődi Ferenc (1953-) (biokémikus) Gonda Sándor (1984-) (gyógyszerész) Vasas Gábor (1975-) (biológus-vegyész) Szücs Boglárka Mikóné Hamvas Márta (1963-) (biológus) Kiss-Szikszai Attila (1975-) (vegyész) Vámosi György (1967-) (biofizikus) Máthé Csaba (1966-) (biológus)
Pályázati támogatás:	NKFIH K120638 OTKA
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM077836
035-os BibID:	(cikkazonosító)2204 (WoS)000447365100115 (Scopus)85052615407
Első szerző:	Gonda Sándor (gyógyszerész)
Cím:	A Simple Method for On-Gel Detection of Myrosinase Activity / Sándor Gonda, Zsolt Szűcs, Tamás Plaszkó, Zoltán Cziáky, Attila Kiss-Szikszai, Gábor Vasas, Márta M-Hamvas
Dátum:	2018
ISSN:	1420-3049
Megjegyzések:	Myrosinase is an enzyme present in many functional foods and spices, particularly in Cruciferous vegetables. It hydrolyses glucosinolates which thereafter rearrange into bioactive volatile constituents (isothiocyanates, nitriles). We aimed to develop a simple reversible method for on-gel detection of myrosinase. Reagent composition and application parameters for native PAGE and SDS-PAGE gels were optimized. The proposed method was successfully applied to detect myrosinase (or sulfatase) on-gel: the detection solution contains methyl red which gives intensive red bands where the HSO 4 ? is enzymatically released from the glucosinolates. Subsequently, myrosinase was successfully distinguished from sulfatase by incubating gel bands in a derivatization solution and examination by LC-ESI-MS: myrosinase produced allyl isothiocyanate (detected in conjugate form) while desulfo-sinigrin was released by sulfatase, as expected. After separation of 80 ?g protein of crude extracts of Cruciferous vegetables, intensive color develops within 10 min. On-gel detection was found to be linear between 0.031?0.25 U (pure Sinapis alba myrosinase, R 2 = 0.997). The method was successfully applied to detection of myrosinase isoenzymes from horseradish, Cruciferous vegetables and endophytic fungi of horseradish as well. The method was shown to be very simple, rapid and efficient. It enables detection and partial characterization of glucosinolate decomposing enzymes without protein purification.
Tárgyszavak:	Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Molecules. - 23 : 9 (2018), p. 1-11. -
További szerzők:	Szűcs Zsolt (1990-) (gyógyszerész) Plaszkó Tamás (1994-) (Biológus Msc) Cziáky Zoltán Kiss-Szikszai Attila (1975-) (vegyész) Vasas Gábor (1975-) (biológus-vegyész) Mikóné Hamvas Márta (1963-) (biológus)
Pályázati támogatás:	NKFIH FK 128021 Egyéb NKFIH PD 124339 Egyéb EFOP-3.6.1-16-2016-00022 EFOP GINOP-2.3.2-15-2016-00008 GINOP
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