Characterization of Ca2+-dependent phospholipase A2 activity during zebrafish embryogenesis.
AuthorsFarber SA, Olson ES, Clark JD, Halpern ME
JournalJ Biol Chem
PubMed ID10383445
'We have developed a simple fluorescent assay for detection of phospholipase A2 (PLA2) activity in zebrafish embryos that utilizes a fluorescent phosphatidylcholine substrate. By using this assay in conjunction with selective PLA2 inhibitors and Western blot analysis, we identified the principal activity in zebrafish embryogenesis as characteristic of the Ca2+-dependent ... More
Down-regulation by elicitors of phosphatidylcholine-hydrolyzing phospholipase C and up-regulation of phospholipase A in plant cells.
AuthorsScherer GF, Paul RU, Holk A, Martinec J
JournalBiochem Biophys Res Commun
PubMed ID12054536
'Phosphatidylcholine, labeled by two fluorescent fatty acids, was fed to cultured plant cells (Petrosilenum crispum, L.; VBI-0, Nicotiana benthiana, L.) and fluorescent diacylglycerol (DAG) was the major metabolite. When a glycoprotein elicitor, derived from Phytophthora sojae, was applied to the parsley cells and the small protein cryptogein from Phytophthora cryptogea ... More
Secretory phospholipase A2 is released from pancreatic beta-cells and stimulates insulin secretion via inhibition of ATP-dependent K+ channels.
AuthorsJuhl K, Efanov AM, Olsen HL, Gromada J
JournalBiochem Biophys Res Commun
PubMed ID14521906
'The release of sPLA(2) from single mouse pancreatic beta-cells was monitored using a fluorescent substrate of the enzyme incorporated in the outer leaflet of the plasma membrane. Stimulation of beta-cells with agents that increased cytosolic free Ca(2+) concentration ([Ca(2+)](i)) induced a rapid release of sPLA(2) to the extracellular medium. Exogenous ... More
Arachidonic acid causes cell death through the mitochondrial permeability transition. Implications for tumor necrosis factor-alpha aopototic signaling.
AuthorsScorrano L, Penzo D, Petronilli V, Pagano F, Bernardi P
JournalJ Biol Chem
PubMed ID11134037
'We have investigated the effects of arachidonic and palmitic acids in isolated rat liver mitochondria and in rat hepatoma MH1C1 cells. We show that both compounds induce the mitochondrial permeability transition (PT). At variance from palmitic acid, however, arachidonic acid causes a PT at concentrations that do not cause PT-independent ... More
Activation by Cdc42 and PIP(2) of Wiskott-Aldrich syndrome protein (WASp) stimulates actin nucleation by Arp2/3 complex.
AuthorsHiggs HN, Pollard TD
JournalJ Cell Biol
PubMed ID10995437
'We purified native WASp (Wiskott-Aldrich Syndrome protein) from bovine thymus and studied its ability to stimulate actin nucleation by Arp2/3 complex. WASp alone is inactive in the presence or absence of 0.5 microM GTP-Cdc42. Phosphatidylinositol 4,5 bisphosphate (PIP(2)) micelles allowed WASp to activate actin nucleation by Arp2/3 complex, and this ... More
Real-time cell assays of phospholipase A(2)s using fluorogenic phospholipids.
AuthorsManna D, Cho W,
JournalMethods Enzymol
PubMed ID17954240
'Phospholipase A(2)s (PLA(2)s) are a superfamily of enzymes involved in production of a wide variety of lipid mediators, including arachidonic acid, lysophospholipids, platelet activation factor, and eicosanoids. Fluorescence-based, real-time cellular activity assays for PLA(2)s have been developed as a tool for studying the function and spatiotemporal regulation of PLA(2)s. Recent ... More
Endocytosis in tobacco pollen tubes: visualisation and measurement of plasma membrane retrieval during different gravity conditions indicates gravity-dependence of endocytosis.
AuthorsLisboa YS, Scherer GE, Quader H
JournalJ Gravit Physiol
PubMed ID15002565
'We studied the effect of microgravity on endocytosis in growing tobacco pollen tubes by measuring the plasma membrane retrieval employing the fluorescent phospholipid bis-Bodipy FL C11- phosphatidylcholine as marker. Time course experiments under 1xg condition revealed a localised and relatively fast plasma membrane retrieval in the pollen tube tip region ... More
Sequence and functional similarities between pro-apoptotic Bid and plant lipid transfer proteins.
AuthorsDegli Esposti M
JournalBiochim Biophys Acta
PubMed ID11997142
'Pro-apoptotic proteins of the Bcl-2 family are known to act on mitochondria and facilitate the release of cytochrome c, but the biochemical mechanism of this action is unknown. Association with mitochondrial membranes is likely to be important in determining the capacity of releasing cytochrome c. The present work provides new ... More
Fluorescent phosphocholine--a specific marker for the endoplasmic reticulum and for lipid droplets in Chara internodal cells.
AuthorsFoissner I,
JournalProtoplasma
PubMed ID19795186
The staining pattern of 1,2-bis(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-undecanoyl)-sn-glycero-3-phosphocholine (Bodipy PC) was investigated in internodal cells of the green alga Chara corallina. Ten minutes after dye addition, Bodipy-PC-derived fluorescence appeared in lipid droplets and after 1 h in the cortical endoplasmic reticulum (ER) and in the inner ER tubes. Staining of the ER required ... More
Fluorogenic substrates for high-throughput measurements of endothelial lipase activity.
AuthorsMitnaul LJ, Tian J, Burton C, Lam MH, Zhu Y, Olson SH, Schneeweis JE, Zuck P, Pandit S, Anderson M, Maletic MM, Waddell ST, Wright SD, Sparrow CP, Lund EG
JournalJ Lipid Res
PubMed ID17090660
Endothelial lipase (EL) has been shown to be a critical determinant for high density lipoprotein cholesterol levels in vivo; therefore, assays that measure EL activity have become important for the discovery of small molecule inhibitors that specifically target EL. Here, we describe fluorescent Bodipy-labeled substrates that can be used in ... More
Elicitor-activated phospholipase A(2) generates lysophosphatidylcholines that mobilize the vacuolar H(+) pool for pH signaling via the activation of Na(+)-dependent proton fluxes.
AuthorsViehweger K, Dordschbal B, Roos W
JournalPlant Cell
PubMed ID12119371
The elicitation of phytoalexin biosynthesis in cultured cells of California poppy involves a shift of cytoplasmic pH via the transient efflux of vacuolar protons. Intracellular effectors of vacuolar proton transport were identified by a novel in situ approach based on the selective permeabilization of the plasma membrane for molecules of ... More
A redox-dependent, G-protein-coupled phospholipase A of the plasma membrane is involved in the elicitation of alkaloid biosynthesis in Eschscholtzia californica.
AuthorsRoos W, Dordschbal B, Steighardt J, Hieke M, Weiss D, Saalbach G
JournalBiochim Biophys Acta
PubMed ID9990291
In cultured cells of California poppy formation of benzophenanthridine alkaloids can be triggered by a yeast elicitor preparation independently of the hypersensitive reaction. A plasma membrane (PM) bound phospholipase A (PLA) is likely to play a role in the signalling process: PLA activity was detectable in individual cells, cell suspensions ... More
A continuous fluorescence assay for the determination of calcium-dependent secretory phospholipase A2 activity in serum.
BACKGROUND: Calcium-dependent secretory phospholipase A(2)-IIA (sPLA(2)-IIA) in the circulation is a marker of inflammation, associated with acute and chronic disease processes. We describe a quick, sensitive and reliable microplate continuous fluorescence assay for determining sPLA(2) activity in serum. METHODS: Liposomes composed of a fluorescent probe and varying amounts of L-alpha-phosphatidylglycerol ... More
Flow cytometric kinetic measurements of neutrophil phospholipase A activation.
AuthorsMeshulam T, Herscovitz H, Casavant D, Bernardo J, Roman R, Haugland RP, Strohmeier GS, Diamond RD, Simons ER
JournalJ Biol Chem
PubMed ID1400459
The interrelationships between activation of phospholipases and neutrophil stimulus-induced Ca2+ responses remain unclear. We report here that immune complexes activate a phosphatidylcholine-specific phospholipase A in a neutrophil only after the cytoplasmic Ca2+ transient has been initiated in the same cell, while chemotactic peptide activation does not proceed via such a ... More
Fluorescence-based assays of lipases, phospholipases, and other lipolytic enzymes.
AuthorsHendrickson HS
JournalAnal Biochem
PubMed ID8059934
In choosing an assay, one needs to consider the following questions: What level of sensitivity is required? Must the assay be continuous? Is the substrate readily available; can it be purchased or must it be custom synthesized? How specific is the substrate? How convenient is the method? How compatible is ... More
Fluorometric assays of phospholipase A2 activity with three different substrates in biological samples of patients with schizophrenia.
AuthorsLasch J, Willhardt I, Kinder D, Sauer H, Smesny S
JournalClin Chem Lab Med
PubMed ID12940517
The rationale of this study was to understand the complexity of kinetics of fluorogenic phospholipid substrates as well as contradictory findings of clinical papers measuring phospholipase A2 (PLA2) activity using different methodologies. The aim was to recommend to clinicians and researchers what substrate in conjunction with what assay should be ... More
Purification and analysis of a phospholipase A2-like lytic factor of Trichomonas vaginalis.
AuthorsLubick KJ, Burgess DE
JournalInfect Immun
PubMed ID14977929
Trichomonas vaginalis produces soluble factors that have been reported to have the ability to damage target cells in vitro, and it has been hypothesized that these factors may play a role in the pathogenesis of human trichomoniasis. A lytic factor (LF) was purified from T. vaginalis, and the molecular characteristics ... More
Identification of a human cDNA clone for lysosomal type Ca2+-independent phospholipase A2 and properties of the expressed protein.
AuthorsKim TS, Sundaresh CS, Feinstein SI, Dodia C, Skach WR, Jain MK, Nagase T, Seki N, Ishikawa K, Nomura N, Fisher AB
JournalJ Biol Chem
PubMed ID8999971
A Ca2+-independent phospholipase A2 (PLA2) maximally active at pH 4 and specifically inhibited by the transition-state analogue 1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol (MJ33) was isolated from rat lungs. The sequence for three internal peptides (35 amino acids) was used to identify a 1653-base pair cDNA clone (HA0683) from a human myeloblast cell line. The ... More
Arachidonic acid released by phospholipase A(2) activation triggers Ca(2+)-dependent apoptosis through the mitochondrial pathway.
AuthorsPenzo D, Petronilli V, Angelin A, Cusan C, Colonna R, Scorrano L, Pagano F, Prato M, Di Lisa F, Bernardi P
JournalJ Biol Chem
PubMed ID15070903
We studied the effects of the divalent cation ionophore A23187 on apoptotic signaling in MH1C1 cells. Addition of A23187 caused a fast rise of cytosolic Ca(2+) ([Ca(2+)](c)), which returned close to the resting level within about 40 s. The [Ca(2+)](c) rise was immediately followed by phospholipid hydrolysis, which could be ... More
U937 cells deprived of endogenous annexin 1 demonstrate an increased PLA2 activity.
AuthorsSolito E, Raguenes-Nicol C, de Coupade C, Bisagni-Faure A, Russo-Marie F
JournalBr J Pharmacol
PubMed ID9756383
Annexin 1 (An 1), a phospholipid and calcium binding protein, is strongly expressed in differentiated U 937 cells. In attempting to correlate the expression of An 1 with phospholipase A2 (PLA2) activity, U 937 cells were stably transfected both with a Sense and Antisense cDNA for An 1. PLA2 activity ... More
Suppression of endothelial or lipoprotein lipase in THP-1 macrophages attenuates proinflammatory cytokine secretion.
Authors
JournalJ Lipid Res
PubMed ID17093291
Involvement of the Saccharomyces cerevisiae hydrolase Ldh1p in lipid homeostasis.
Authors
JournalEukaryot Cell
PubMed ID21478434
Genetic and structure-function studies of missense mutations in human endothelial lipase.
Authors
JournalPLoS One
PubMed ID23536757
The lipid transporter Mfsd2a maintains pulmonary surfactant homeostasis.
Authors
JournalJ Biol Chem
PubMed ID35150739
TRPV4 channel opening mediates pressure-induced pancreatitis initiated by Piezo1 activation.