ArticleJournal of lipid research2009
Enzymatic measurement of phosphatidic acid in cultured cells.
Article in Journal of lipid research, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 51 citations in OpenAlex.
- Phospholipid biomarkers of coronary heart disease.Journal of pharmaceutical health care and sciences · 2024Review
- Acting as a Molecular Tailor: Dye Structural Modifications for Improved Sensitivity toward Lysophosphatidic Acids Sensing.ACS omega · 2023Article
- A Peptide-Based Fluorescent Sensor for Anionic Phospholipids.ACS omega · 2022Article
- Alterations in cellular and organellar phospholipid compositions of HepG2 cells during cell growth.Scientific reports · 2021Article
- Nuclear lipid droplets form in the inner nuclear membrane in a seipin-independent manner.The Journal of cell biology · 2021Article
- Protocols for Enzymatic Fluorometric Assays to Quantify Phospholipid Classes.International journal of molecular sciences · 2020Review
- Classical Activation of Macrophages Leads to Lipid Droplet Formation WithoutFrontiers in immunology · 2020Article
- Lipid class composition of membrane and raft fractions from brains of individuals with Alzheimer's disease.Biochemistry and biophysics reports · 2019Article
- Dopey1-Mon2 complex binds to dual-lipids and recruits kinesin-1 for membrane trafficking.Nature communications · 2019Article
- Enzymatic fluorometric assays for quantifying all major phospholipid classes in cells and intracellular organelles.Scientific reports · 2019Article
- Attenuation of liver cancer development by oral glycerol supplementation in the rat.European journal of nutrition · 2018Article
- Normal human adipose tissue functions and differentiation in patients with biallelicJournal of lipid research · 2017Article
- YeastThe Journal of biological chemistry · 2017Article
- Tips on the analysis of phosphatidic acid by the fluorometric coupled enzyme assay.Analytical biochemistry · 2017Article
- Cholesterol attenuates cytoprotective effects of phosphatidylcholine against bile salts.Scientific reports · 2017Article
- Phosphorylation of Dgk1 Diacylglycerol Kinase by Casein Kinase II Regulates Phosphatidic Acid Production in Saccharomyces cerevisiae.The Journal of biological chemistry · 2016Article
- Enzymatic measurement of phosphatidylglycerol and cardiolipin in cultured cells and mitochondria.Scientific reports · 2015Article
- Intramembrane proteolysis of β-amyloid precursor protein by γ-secretase is an unusually slow process.Biophysical journal · 2015Article
- Distinct roles of the phosphatidate phosphatases lipin 1 and 2 during adipogenesis and lipid droplet biogenesis in 3T3-L1 cells.The Journal of biological chemistry · 2013Article
- Diacylglycerol kinases terminate diacylglycerol signaling during the respiratory burst leading to heterogeneous phagosomal NADPH oxidase activation.The Journal of biological chemistry · 2013Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In this work, we developed a novel enzymatic method for measuring phosphatidic acid (PA) in cultured cells. The enzymatic reaction sequence of the method involves hydrolysis of PA to produce glycerol-3-phosphate (G3P), which is then oxidized by G3P oxidase to generate hydrogen peroxide. In the presence of peroxidase, hydrogen peroxide reacted with Amplex Red to produce highly fluorescent resorufin. We found that lipase from Pseudomonas sp. can completely hydrolyze PA to G3P and FAs. The calibration curve for PA measurement was linear between 20 and 250 microM, and the detection limit was 5 microM (50 pmol in the reaction mixture). We also modified the method for the enzymatic measurement of lysophosphatidic acid. By this new method, we determined the PA content in the lipid extract from HEK293 cells. The cellular content of PA was decreased with increasing cell density but not correlated with the proliferation rate. The diacylglycerol kinase inhibitor R59949 markedly reduced the cellular PA content, suggesting the diacylglycerol kinase activity was involved in a large part of the PA production in HEK293 cells. This novel method for PA quantification is simple, rapid, specific, sensitive, and high-throughput and will help to study the biological functions of PA and its related enzymes.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.