Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.4.3 (phospholipase C)
 18,461 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We report here the inhibition of enzyme activity of phospholipase C-anti-human rabbit IgG conjugate complexed with human IgG. Phospholipase C activity was measured by assaying the release of hemoglobin from erythrocytes. The degree of inhibition varies depending on the relative amount of IgG utilized. A water soluble carbodiimide linking yields a conjugate which is inhibited by IgG, while conjugate prepared by glutaraldehyde is not. Immunodiagnostic assays requiring detection of low levels of antigen or antibody may be possible utilizing the phospholipase C-erythrocyte technique.
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PMID:Phospholipase C-labeled anti-human IgG: inhibition by human IgF. 32 66

Erythrocytes from several different species were exposed to Ca2+ and the bivalent-cation ionophore A23187. The lipid composition, morphology and K+ permeability of the treated cells were investigated. Erythrocytes from human, rat, guinea pig and rabbit (a) showed an increased concentration of 1,2-diacyl-sn-glycerol and enhanced labelling of phosphatidate with 32P, (b) underwent echinocytosis and outward vesiculation, and (c) rapidly released much of their intracellular K+. Pig cells showed only the K+ loss, and ox and sheep (high-K+) cells showed none of these Ca2+-evoked effects. All of the cells underwent stomatocytosis and inward vesiculation when treated externally with Clostridium perfringens phospholipase C. These results support the idea that there is a correlation between the asymmetric insertion of diacylglycerol (or ceramide) into the membrane and the shape-changes leading to microvesiculation, but they indicate that Ca2+-triggered K+ efflux and diacylglycerol production are unrelated events. Erythrocytes of chicken and turkey showed no Ca2+-stimulated K+ efflux. They showed slight ionophore A23187-stimulated vesiculation, but this appeared to be associated with the appearance in the membrane of ceramide rather than of diacylglycerol. Phospholipase C treatment caused very similar changes in morphology and phosphatidate labelling to those seen in mammalian erythrocytes.
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PMID:Calcium ion-dependent diacylglycerol accumulation in erythrocytes is associated with microvesiculation but not with efflux of potassium ions. 33 8

1. Phospholipase C was inactivated by exposure to the three amino-group reagents, ethyl acetamidate, 2,4,6-trinitrobenzensulphonic acid and pyridoxal 5'-phosphate plus reduction. 2. Inactivation by pyridoxal 5'-phosphate showed the characteristics of Schiff's base formation with the enzyme. The pyridoxal 5'-phosphate-treated enzyme after reduction had an absorbance maximum at 325 mm and 6-N-pyridoxyl-lysine was the only fluorescent component after acid hydrolysis. 3. For complete inactivation, 2 mol of pyridoxal 5'-phosphate or 7 mol of 2,4,6-trinitrophenyl were incorporated/mol of enzyme. 4. The two apparently essential lysine residues were much more reactive to pyridoxal 5'-phosphate than the other 19 lysine residues in the enzyme. 5. Binding of phospholipase C to a substrate-based affinity gel caused marked protection against inactivation by pyridoxal 5'-phosphate. For complete inactivation of the gel-bound enzyme, 5 mol of pyridoxal 5'-phosphate were incorporated/mol of enzyme and there was no evidence of two especially reactive lysine residues. 6. On application of pyridoxal 5'-phosphate-treated enzyme (remaining activity 30% of original) to a column of the affinity gel, some material bound and some did not. The latter contained very little enzyme activity and was heavily incorporated with reagent (9.06 mol/mol of enzyme). The former had a specific activity of 34% of that of the control and contained 1.29 mol of reagent/mol of enzyme. 7. Thus phospholipase C appears to contain two lysine residues that are essential for enzyme activity, but probably not for substrate binding.
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PMID:Phospholipase C from Bacillus cereus. Evidence for essential lysine residues. 40 7

Phospholipase C from Bacillus cereus was inactivated by incubation with either of the carboxyl reagents, a water-soluble carbodimide plus a nucleophile or Woodward's reagent K. With the former reagent, the incorporation into the enzyme of the first mol of nucleophile caused a 4-5-fold increase in the Km for dihexanoyllecithin with no significant effect on the Vm. The second mol of nucleophile incorporated caused no further change in Km but destroyed most of the catalytic activity. Modification of the enzyme by carbodiimide plus nucleophile did not alter the relative activity of the enzyme towards micelles and monomolecularly dispersed solutions of diheptanoyllecithin. Furthermore, inactivation by this reagent did not significantly decrease the ability of the enzyme to bind to a substrate-based affinity gel. It was concluded that phospholipase C contains a single carboxyl group that is essential for catalytic activity. The enzyme also contains a total of 4-5 reactive/exposed carboxyl groups.
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PMID:Inactivation of phospholipase C from Bacillus cereus by a carboxyl group modifying reagent. 40 15

The effect of modification of photoreceptor membranes of the bovine retina on the termodynamical parameters that characterize heat denaturation of rodopsin was studied. The highest increase of the rate constant and the corresponding maximal drop of the free energy change of heat denaturation of the pigment were obtained by using 7 M urea or 25% Triton X-100 in the presence of 5.10(-4) M EDTA. After chipping off one third of the protein from the rodopsin molecule by papain treatment a significant decrease of the slope of the Arrenius curve and a maximal decrease of entropy change compared to the parameters known for heat denaturation of the pigment in native photoreceptor membranes were found. Modification of the lipid components of the photoreceptor membranes (treatment with Triton X-100 and phospholipase C) reduced the thermostability of rodopsin. Maximal changes were obtained at Triton X-100 concentrations 0.1--1%, further concentration increas (1--25%) did not lead to significant changes. Phospholipase C treatment resulted in a decrease of free energy change and an increase of entropy change without affecting entalpy changes, accompaning the heat denaturation of rodopsin. Bivalent cations (Ca2+, Mg2+) increased the termostability of rodopsin both in photoreceptor membranes and in solutions to 25% Triton X-100.
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PMID:[Modification of the retina photoreceptor membranes and temperature stability of rhodopsin]. 73 88

The effect of phospholipase C (EC 3.1.4.3) on human blood platelets has been studied. Phospholipase C from Bacillus cereus was purified to homogeneity as judged by analytical and sodium dodecyl sulphate disc gel electrophoresis and by immunoelectrophoresis. Human platelets isolated from platelet-rich plasma by gel filtration or by centrifugation and washing were incubated with phospholipase C. A loss of 20-45% of the total platelet phospholipid was observed, whereas 88% was hydrolyzed when platelet homogenates were submitted to identical enzyme treatment. Intact platelets lost 50-75% phosphatidylethanolamine, 20-50% phosphatidylcholine, and 20-25% phosphatidylserine. Sphingomyelin was not a substrate for the enzyme under the conditions used. The platelets contained no detectable endogenous phospholipase C activity. The loss of phospholipid was not accompanied by aggregation of the platelets, nor did the platelets lose their ability to aggregate with ADP or thrombin. Total platelet factor 3 releasable by freezing and thawing was reduced. Measurements of releasable platelet factor 4 and the efflux of serotonin showed that no release reaction was triggered even when up to 45% of the total phospholipid in the platelets was hydrolyzed. When sphingomyelinase was added together with, before, or after phospholipase C, aggregation occurred. Sphingomyelinase alone gave no aggregation. The gel-filtered platelets also aggregated upon addition of purified phospholipase C from Clostridium perfringens. The distribution of phospholipids in the platelet membrane is discussed.
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PMID:The effect of phospholipase C on human blood platelets. 81 57

Phospholipase C [EC 3.1.4.3] from Pseudomonas aureofaciens was found to be inhibited by chelating reagents such as ethylenediaminetetraacetate [EDTA] and o-phenanthroline. The inhibition was reversed by the addition of Zn2+ and, to a lesser extent, by Co2+ and Mn2+. On isoelectric focusing, the isoelectric point of this enzyme proved to be 6.3--6.5, with a single peak. The enzyme reaction with the substrate was followed in media containing an organic solvent such as diethyl ether or diethyl ether-ethyl alcohol. When ethyl alcohol was added (up to 2%) to the reaction mixture in ether, there were no marked changes in the hydrolytic rates of phosphatidylcholine and phosphatidylethanolamine. However, the enzyme activity was inhibited when the alcohol concentration was increased above 2%. In 98% diethyl ether-2% ethyl alcohol, phosphatidylcholine was hydrolyzed more rapidly than phosphatidylethanolamine, in contrast with the result obtained in water. In the single micelle state, phosphatidylethanolamine was hydrolyzed more rapidly than phosphatidylcholine or lysophatidylcholine. Acidic phospholipids and sphinogomyelin were not hydrolyzed. When the enzyme was incubated with phospholipid mixture extracted from Ps aureofaciens and rat liver, both phosphatidylethanolamine and phosphatidylcholine were hydrolyzed more rapidly than in the single micelle state of these substrates.
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PMID:Studies on phospholipase C from Pseudomonas aureofaciens. II. Further studies on the properties of the enzyme. 82 22

1. The effects of phospholipases A from bee venom and from porcine pancreas and of phospholipases C from Clostridium welchii and Bacillus cereus on active and passive membrane properties of Aplysia neurones have been studied. Consistent alterations in electrical membrane properties were found following intracellular application of three of these enzymes.2. Bee venom phospholipase A produced a rapid decrease of membrane potential and resistance. Voltage clamping revealed a marked depression of peak transient current with little or no effect in the late outward current.3. Mammalian phospholipase A was found ineffective in changing either the resting or active membrane properties.4. Phospholipase C from Bacillus cereus led to a strong hyperpolarization and a fall in membrane resistance. Voltage clamping revealed a marked increase in the late outward current.5. Neurones injected with Clostridium welchii phospholipase C manifested a several-fold rise in resting membrane resistance as well as a tendency to slight hyperpolarization.6. All enzymes were ineffective when externally applied.7. It is tentatively concluded that the internally applied phospholipases affect specific ionic permeabilities both in the resting and active excitable membrane. Various mechanisms by which the differing actions of enzymes of the same type could be explained are discussed.
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PMID:Membrane properties of Aplysia neurones intracellularly injected with phospholipases A and C. 87 92

The role of phospholipids in the activity of UDP-D-galactose: D-xylose galactosyltransferase (galactosyltransferase I) from embryonic chick cartilage was investigated. Phospholipase C treatment of particulate galactosyltransferase I caused inactivation of this enzyme to the extent of 60 to 70% as well as hydrolysis of 75 to 80% of the membrane phospholipids. Addition of phospholipid restored activity to nearly control levels. The order of effectiveness of various phopholipids in reactivating phospholipase C-treated galactosyltransferase I was as follows: lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidylethanolamine, phosphatidylcholine. The effect of phospholipase A on galactosyltransferase I activity was also examined and was found to be concentration-dependent. At concentrations less than 10 mug/mg of pellet protein, phospholipase A slightly activated galactosyltransferase I. whereas at higher concentrations it inhibited the activity in a manner similar to phospholipase C. Galactosyltransferase I was activated moderately and also solubilized by treatment with Nonidet P-40 in the presence of 0.5 M KCl. Following solubilization and purification by gel filtration and affinity chromatography, galactosyltransferase I could be inactivated by detergent removal by dialysis and subsequently reactivated by addition of detergent. Neither phospholipase C treatment nor exogenous phospholipid had any significant effect on three of the other chondroitin sulfate glycosyltransferases (UDP-D-xylose: core protein xylosyltransferase, UDP-D-glucuronic acid:3-O-beta-D-galactosyl-D-galactose glucuronosyltransferase, and UDP-N-acetyl-D-galactosamine:(BlcUA-GalNAc-4-sulfate)j N-acetylgalactosaminyltransferase). On lipid analysis by thin layer chromatography, phosphatidylcholine and phosphatidylethanolamine were found to be the major phospholipids of particulate and solubilized glycosyltransferase preparations from embryonic chick cartilage, while lysopholipids of particulate and solubilized glycosyltransferase preparations from embryonic chick cartilage, while lysophosphatidylcholine and lysophosphatidylethanolamine were barely detectable components. The concentration of these specific phospholipids was diminished greatly following phospholipase C treatment.
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PMID:Biosynthesis of chondroitin sulfate. Role of phospholipids in the activity of UDP-D-galactose: D-xylose galactosyltransferase. 94 16

Infusions of tissue thromboplastin induce intravascular coagulation in animals. Phospholipase C (PLC) (EC 3.1.4.3) from Bacillus cereus has a marked protective effect against such infusions and might be of value in the therapy of certain types of intravascular coagulation. We have therefore studied the toxicity, half-life, and effect on lipolysis of purified PLC in rats, using parenteral administration of 14C-labelled enzyme. Following intravenous injection, the plasma half-life was 5.2-5.4 min, and LD50 was approximately 1.6-1.7 mg/kg. The effect of PLC on lipolysis was moderate. The enzyme does not appear to be bound to any plasma macromolecules, and there was no accumulation of labelled enzyme in tissues other than kidney.
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PMID:Parenteral administration of phospholipase C in the rat. Distribution, elimination, and lethal doses. 100 44


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