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Statements

Subject Item
n2:RIV%2F61389030%3A_____%2F13%3A00423956%21RIV14-GA0-61389030
rdf:type
n16:Vysledek skos:Concept
dcterms:description
Plant nonspecific phospholipase C (NPC) is a recently described enzyme which plays a role in membrane rearrangement during phosphate starvation. It is also involved in responses of plants to brassinolide, abscisic acid (ABA), elicitors, and salt. The NPC activity is decreased in cells treated with aluminum. In the case of salt stress, the molecular mechanism of NPC action is based on accumulation of diacylglycerol (DAG) by hydrolysis of phospholipids and conversion of DAG, the product of NPC activity, to phosphatidic acid (PA) that participates in ABA signaling pathways. Here we describe a step-by-step protocol, which can be used to determine in situ or in vitro NPC activity. Determination is based on quantification of fluorescently labeled DAG as a product of cleavage of the fluorescently labeled substrate lipid, phosphatidylcholine. High-performance thin-layer chromatography is used for separation of fluorescent DAG. The spot is visualized with a laser scanner and the relative amounts of fluorescent DAG are quantified using imaging software. Plant nonspecific phospholipase C (NPC) is a recently described enzyme which plays a role in membrane rearrangement during phosphate starvation. It is also involved in responses of plants to brassinolide, abscisic acid (ABA), elicitors, and salt. The NPC activity is decreased in cells treated with aluminum. In the case of salt stress, the molecular mechanism of NPC action is based on accumulation of diacylglycerol (DAG) by hydrolysis of phospholipids and conversion of DAG, the product of NPC activity, to phosphatidic acid (PA) that participates in ABA signaling pathways. Here we describe a step-by-step protocol, which can be used to determine in situ or in vitro NPC activity. Determination is based on quantification of fluorescently labeled DAG as a product of cleavage of the fluorescently labeled substrate lipid, phosphatidylcholine. High-performance thin-layer chromatography is used for separation of fluorescent DAG. The spot is visualized with a laser scanner and the relative amounts of fluorescent DAG are quantified using imaging software.
dcterms:title
Assaying nonspecific phospholipase C activity Assaying nonspecific phospholipase C activity
skos:prefLabel
Assaying nonspecific phospholipase C activity Assaying nonspecific phospholipase C activity
skos:notation
RIV/61389030:_____/13:00423956!RIV14-GA0-61389030
n16:predkladatel
n17:ico%3A61389030
n3:aktivita
n5:Z n5:P
n3:aktivity
P(GAP501/12/1942), P(GPP501/12/P950), Z(AV0Z50380511)
n3:dodaniDat
n4:2014
n3:domaciTvurceVysledku
n7:9481923 n7:9878572
n3:druhVysledku
n13:C
n3:duvernostUdaju
n19:S
n3:entitaPredkladatele
n22:predkladatel
n3:idSjednocenehoVysledku
62213
n3:idVysledku
RIV/61389030:_____/13:00423956
n3:jazykVysledku
n10:eng
n3:klicovaSlova
Bodipy; Diacylglycerol; Nonspecific phospholipase C
n3:klicoveSlovo
n9:Diacylglycerol n9:Bodipy n9:Nonspecific%20phospholipase%20C
n3:kontrolniKodProRIV
[D80BF93F3F2C]
n3:mistoVydani
New York City
n3:nazevZdroje
Plant Lipid Signaling Protocols
n3:obor
n15:ED
n3:pocetDomacichTvurcuVysledku
2
n3:pocetStranKnihy
305
n3:pocetTvurcuVysledku
3
n3:projekt
n12:GAP501%2F12%2F1942 n12:GPP501%2F12%2FP950
n3:rokUplatneniVysledku
n4:2013
n3:tvurceVysledku
Pejchar, Přemysl Martinec, Jan Günther, F.E.S.
n3:zamer
n14:AV0Z50380511
s:numberOfPages
11
n18:doi
10.1007/978-1-62703-401-2_18
n20:hasPublisher
Humana Press
n11:isbn
978-1-62703-400-5