A microplate technique for phospholipase D activity determination

Sajdok, J.; Jandus, J.; Valentová, Olga; Novotná, Zuzana; Káš, Jan; Daussant, J.

doi:10.1016/0003-2670(95)00308-m

Cited by 17 publications

(13 citation statements)

References 5 publications

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“…PLDα activity was assayed spectrophotometrically as described by Sajdok et al. (1995), and PIP 2 ‐dependent PLD (PLDβ/γ) activity was determined radiometrically as described by Pappan et al.…”

Section: Methodsmentioning

confidence: 99%

Mutual regulation of plant phospholipase D and the actin cytoskeleton

et al. 2010

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Summary Membrane lipids and cytoskeleton dynamics are intimately inter‐connected in the eukaryotic cell; however, only recently have the molecular mechanisms operating at this interface in plant cells been addressed experimentally. Phospholipase D (PLD) and its product phosphatidic acid (PA) were discovered to be important regulators in the membrane–cytoskeleton interface in eukaryotes. Here we report the mechanistic details of plant PLD–actin interactions. Inhibition of PLD by n‐butanol compromises pollen tube actin, and PA rescues the detrimental effect of n‐butanol on F‐actin, showing clearly the importance of the PLD–PA interaction for pollen tube F‐actin dynamics. From various candidate tobacco PLDs isoforms, we identified NtPLDβ1 as a regulatory partner of actin, by both activity and in vitro interaction assays. Similarly to published data, the activity of tobacco PIP2‐dependent PLD (PLDβ) is specifically enhanced by F‐actin and inhibited by G‐actin. We then identified the NtPLDβ1 domain responsible for actin interactions. Using sequence‐ and structure‐based analysis, together with site‐directed mutagenesis, we identified Asn323 and Thr382 of NtPLDβ1 as the crucial amino acids in the actin‐interacting fold. The effect of antisense‐mediated suppression of NtPLDβ1 or NtPLDδ on pollen tube F‐actin dynamics shows that NtPLDβ1 is the active partner in PLD–actin interplay. The positive feedback loop created by activation of PLDβ by F‐actin and of F‐actin by PA provides an important mechanism to locally increase membrane–F‐actin dynamics in the cortex of plant cells.

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Section: Methodsmentioning

confidence: 99%

Mutual regulation of plant phospholipase D and the actin cytoskeleton

et al. 2010

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“…2. ATX activity, according to Sajdok et al [15], is based on colorimetric determination of the product of the ATX reaction, choline, using choline oxidase/ peroxidase system, by the microtitration plate technique at 490 nm using an ELISA reader. The following reagents were used: choline oxidase (Sigma- M (MW 288.4) equals to 3 mg; 9% Triton X-100 was prepared by diluting 9 mL of Triton X-100 in 100 mL with distilled water; potassium phosphate buffer (0.2 M, pH 6); 5% SDS was prepared by dissolving 0.5 g of SDS in 10 mL of distilled water; a termination mixture of ATX reaction: 50 mM (EDTA) (MW 292.…”

Section: I-serummentioning

confidence: 99%

Effect of histidine on autotaxin activity in experimentally induced liver fibrosis

El-Batch

Ibrahim

Said

2010

J Biochem & Molecular Tox

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The aim of this study was to explain whether serum autotaxin (ATX) activity might be a target for regulation of liver fibrosis and to evaluate the hepatoprotective and antifibrotic effects of histidine in thioacetamide (TAA)-induced liver fibrosis in rats. This study was carried out on 100 Wistar Albino rats, classified into five groups, each containing 20 rats: Group I (control group), Group II: rats were given histidine intraperitoneally, Group III: rats were injected intraperitoneally with TAA, Group IV: rats were injected with L-histidine together with TAA, and Group V: rats were injected with TAA for 1 month then treated with intraperitoneal injection of L-histidine for another month. At the end of experiment, blood and liver were collected for determination of some liver enzymes, plasma total antioxidant capacity (TAC), serum ATX activity, and liver tissue hydroxyproline. Thioacetamide treatment caused significant increases in liver enzymes, ATX activities, and liver hydroxyproline, but a significant decrease in plasma's TAC. Upon treatment with histidine, a significant decrease in liver enzymes, ATX activities, and liver hydroxyproline was observed with a significant increase in plasma TAC in Group IV and a significant decrease in Group V. Histidine as an antioxidant has a protective effect on TAA-induced liver fibrosis; it is beneficial in rats not only by inhibition of collagen synthesis and increasing TAC but also by inhibition of ATX activities thus reducing its capacity to produce lysophosphatidic acid, which has a role in liver fibrosis. C 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 25:143-150, 2011; View this article online at wileyonlinelibrary.com.

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“…Hydrolytic PIP 2 -independent PLD activity was assayed by the spectrophotometric method described by Sajdok et al (1995) in the presence of PC as a substrate, 120 mM Ca 2+ and 10 mM SDS in the reaction mixture.…”

Section: Assay Of Pld Hydrolytic Activitymentioning

confidence: 99%

Phosphatidic acid produced by phospholipase D is required for tobacco pollen tube growth

Potocký

Eliáš

Profotová

et al. 2003

Planta

161

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Phospholipase D (PLD) and its product phosphatidic acid (PA) are involved in a number of signalling pathways regulating cell proliferation, membrane vesicle trafficking and defence responses in eukaryotic cells. Here we report that PLD and PA have a role in the process of polarised plant cell expansion as represented by pollen tube growth. Both phosphatidylinositol-4,5-bisphosphate-dependent and independent PLD activities were identified in pollen tube extracts, and activity levels during pollen tube germination and growth were measured. PLD-mediated PA production in vivo can be blocked by primary alcohols, which serve as a substrate for the transphosphatidylation reaction. Both pollen germination and tube growth are stopped in the presence 0.5% 1-butanol, whereas secondary and tertiary isomers do not show any effect. This inhibition could be overcome by addition of exogenous PA-containing liposomes. In the absence of n-butanol, addition of a micromolar concentration of PA specifically stimulates pollen germination and tube elongation. Furthermore, a recently established link between PLD and microtubule dynamics was supported by taxol-mediated partial rescue of the 1-butanol-inhibited pollen tubes. The potential signalling role for PLD-derived PA in plant cell expansion is discussed.

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A microplate technique for phospholipase D activity determination

Cited by 17 publications

References 5 publications

Mutual regulation of plant phospholipase D and the actin cytoskeleton

Mutual regulation of plant phospholipase D and the actin cytoskeleton

Effect of histidine on autotaxin activity in experimentally induced liver fibrosis

Phosphatidic acid produced by phospholipase D is required for tobacco pollen tube growth

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