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Biomedical subjects

G Graff

Publications and source records attributed to G Graff.

At least 55 records · Page 3Linked to original sources
PubMed1981

Cyclic GMP metabolism in psoriasis: increased activity of soluble epidermal cyclic GMP phosphodiesterase and its modulation by calcium.

Soluble cyclic GMP-phosphodiesterase was measured in normal and psoriatic epidermis. The specific activity of the enzyme was increased almost four-fold in involved compared with normal epidermis, and two- to three-fold in involved compared with uninvolved epidermis. The enzyme activity from all three sources was inhibited by 40-50% by ethylene glycol tetraacetic acid (EGTA). These results indicate that in addition to the reported enhanced capacity of psoriatic epidermis to generate cGMP, it has an increased ability to hydrolyse this nucleotide, although to a lesser degree than the augmentation found in soluble guanylate activity from psoriatic epidermis. These observations are compatible with the elevated steady-state levels of this nucleotide observed in the involved epidermis of psoriasis.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1980

18O-Labeling of guanosine monophosphate upon hydrolysis of cyclic guanosine 3':5'-monophosphate by phosphodiesterase.

The hydrolysis of cGMP by phosphodiesterase was conducted in [18O]water to determine the site of bond cleavage and the stoichiometry of 18O incorporation into 5'-GMP. Three different forms of phosphodiesterase including a calmodulin-calcium-dependent enzyme in its basal and activated states were examined. The hydrolysis of cGMP catalyzed by each of the forms of phosphodiesterase proceeded with incorporation of 1 18O atom recoverable in the phosphate moiety of each molecule of 5'-GMP generated. No molecular species of phosphate deriving from the 5'-GMP generated containing two or three 18O were detectable. These results indicate that the phosphodiesterase-catalyzed hydrolysis of cGMP proceeds by nucleophilic substitution at phosphorus resulting in P-O bond cleavage. The stoichiometry of 18O incorporation indicates that the reaction proceeds without phosphate-water oxygen exchange when the hydrolytic reaction is catalyzed by diverse forms of phosphodiesterase in the basal or activated state. These considerations of the phosphodiesterase reaction help to establish the validity of monitoring the rate of enzyme-catalyzed hydrolysis of cGMP as a function of the rate of 18O-labeling of the phosphate of 5'-GMP when the reaction proceeds in a medium of predetermined 18O enrichment.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1980

Human placental oxytocinase and its relationship to pregnancy plasma oxytocinase.

Plasma "oxytocinase of pregnancy" and three placental "oxytocinase" fractions from human placental extracts were compared. On the basis of acrylamide-agarose chromatography, polyacrylamide gel electrophoresis, substrate specificity, heat liability and relative insensitiveness to L-methionine, it is concluded that placental enzyme activities, present in the second acrylamide-agarose peak, are identical with the plasma "pregnancy oxytocinase" and are to be regarded as its source. The hypothesis of a transplacental passage of placental peak II oxytocinase in the blood compartment of the mother seems well supported. Heat treatment of the placental extracts uncovered a minor activity behaving like Oya's microsomal oxytocinase, which is totally unlike the plasma oxytocinase and plays no part in the increased oxytocinase activity of human pregnancy plasma. A hitherto undescribed enzyme activity shares with the pregnancy oxytocinase its specificity towards L-leucyl-beta-naphthylamide and di-S-S-L-cysteinyl-beta-naphthylamide, its heat lability and realtive insensitiveness to L-methionine. However, this activity is carried by a protein of much lower molecular weight as judged by acrylamide-agarose chromatography, which shows only a single activity band on polyacrylamide gel electrophoresis.

Acid Phosphatase↗
PubMed1980

1-arachidonyl-monoglyceride causes platelet aggregation: indirect evidence for an acylglycerol acylhydrolase involvement in the release of arachidonic acid for prostaglandin synthesis.

Phosphatidylcholine liposomes containing 1-arachidonyl-monoglyceride were found to cause aggregation of human platelets. In contrast, addition of phosphatidylcholine liposomes, 1-arachidonyl-monoglyceride, or phosphatidylcholine liposomes containing I-oleoyl-monoglyceride to a similar platelet preparation had no effect. Aggregation stimulated by 1-arachidonyl-monoglyceride was inhibited by 100 microM aspirin or 1 microM indomethacin, suggesting that the arachidonic acid is first released by; a platelet acylglycerol acylhydrolase and then converted to PGG2 and thromboxane A2 which initiate the platelet aggregation. Changes in platelet morphology in response to 1-arachidonyl-monoglyceride were similar to those reported previously to occur following stimulation of platelets by arachidonic acid or PGG2 providing further support for this concept. EDTA inhibited aggregation of platelets but no shape change or granule centralization in response to 1-arach-idonyl-monoglyceride. PGE1 and theophylline inhibited both aggregation and morphological changes. These results with inhibitors are similar to the effects of these inhibitors on PGG2 and provide further evidence for similarity between the action of 1-arachidonyl-monoglyceride and PGG2. The results provide important evidence to support the concept that an acylglycerol acylhydrolase may be involved in arachidonic acid release and platelet aggregation.

Arachidonic Acids↗
PubMed1980

Cyclic GMP metabolism in psoriasis: activation of soluble epidermal guanylate cyclase by arachidonic acid and 12-hydroxy-5,8,10,14-eicosatetraenoic acid.

Soluble guanylate cyclase activity was measured in normal and psoriatic human epidermis. The specific activity of guanylate cyclase was determined to be increased 10-fold and 3-fold in involved and uninvolved epidermis of psoriatics, respectively, compared to normal epidermis. Arachidonic acid (5 to 100 micrometers) or 12-hydroxy-5,8,10,14-eicosatetraenoic acid (HETE) (5 to 50 micrometers) stimulated guanylate cyclase activity from involved epidermis 2- to 3-fold and from uninvolved epidermis up to 2-fold, but these fatty acids had no effect on the activity of this cyclase from normal epidermis. These results indicate that there is an increase in the cGMP biosynthetic capacity of involved epidermis from psoriatics that derives from a markedly increased specific activity of guanylate cyclase and an alteration in a property of this enzyme activity which renders it responsive to fatty acids reported to accumulate in this lesion. These observations are consistent with the report that an elevated steady-state level of cGMP is one of the consequences of the strikingly altered metabolism of cGMP in psoriatic epidermis.

Arachidonic Acids↗
PubMed1980

Increased activity and stimulability of psoriatic epidermal soluble guanylate cyclase by arachidonic acid and 12-hydroxy-5,8,10,14-eicosatetraenoic acid.

Soluble guanylate cyclase activity was measured in normal and psoriatic human epidermis. The specific activity of guanylate cyclase was determined to be increased 10-fold and 3-fold in involved and uninvolved epidermis of psoriatics, respectively, compared with normal epidermis. Arachidonic acid or HETE (5 to 50 microM) stimulated guanylate cyclase activity from involved epidermis 2 to 3-fold and from uninvolved epidermis up to 2-fold, but these fatty acids had no effect on the activity of this cyclase from normal epidermis. These results indicate that the biosynthetic capacity to generate cGMP is markedly increased in involved epidermis from psoriatics because of a markedly increased specific activity of guanylate cyclase and an alteration in a property of this enzyme activity which renders it stimulable by fatty acids reported to accumulate in this lesion.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1979

Identification of 15-keto-9, 11-peroxidoprosta-5, 13- dienoic acid as a hematin-catalyzed decomposition product of 15-hydroperoxy-9, 11-peroxidoprosta-5, 13-dienoic acid.

A labile prostaglandin was isolated as one of the products generated from [1-14C] eicosatetraenoic acid incubated with sheep vesicular gland microsomes. The eicosatetraenoic acid metabolite amounted to ca. 16% of the total radiolabeled products. Formation of this new prostaglandin was prevented when heat-denatured microsomes were employed or when incubation mixtures were supplemented with indomethacin or phenol. However, incubation of prostaglandin G2 (PGG2) with hematin in the presence or absence of catalytically active or heat-inactivated microsomes led to production of approximately the same quantity of the new prostaglandin. These results indicated that the new prostaglandin can be formed nonenzymically. The new prostaglandin was conclusively identified by gas liquid chromatography-mass spectrometry analysis as 15-keto-9,11-peroxidoprosta-5,13-dienoic acid (15-keto-PGG2) after chemical conversion to known prostaglandins. The effects of 15-keto-PGG2 and PGG2 were similar on canine lateral saphenous vein; both promoted contraction followed by prolonged relaxation, but 15-keto-PGG2 appeared to be 1/50 as potent as PGG2.

Animals↗
PubMed1978

Activation of soluble splenic cell guanylate cyclase by prostaglandin endoperoxides and fatty acid hydroperoxides.

Purified prostaglandin endoperoxides (PGG2 and PGH2) and hydroperoxides (15-OOH-PGE2) as well as fatty acid hydroperoxides (12-OOH-20:4, 15-00H-20:4, and 13-OOH-18:2) were examined as effectors of soluble splenic cell guanylate cyclase activity. The procedures described (in the miniprint supplement) for the preparation, purification, and characterization of these components circumvented the use of diethyl ether which obscured effects of lipid effectors because of contaminants presumed to be ether peroxides which were stimulatory to the cyclase. Addition of prostaglandin endoperoxides or fatty acid hydroperoxides to the reaction mixture led to a time-dependent activation of guanylate cyclase activity; 2.5- to 5-fold stimulation was seen during the first 6 min. The degree of stimulation and rate of activation were dependent on the concentration of the fatty acid effector; when initial velocities (6 min) were assessed half-maximal stimulation was achieved in the range of 2 to 3 micrometer. However, by extending the incubation time to 90 min similar maximal increases in specific activity could be achieved with 3 or 10 micrometer PGG2 or PGH2. Activation of guanylate cyclase upon addition of prostaglandin endoperoxides or fatty acid hydroperoxides was prevented or reversed by the thiol reductants dithiothreitol (3 to 5 mM) or glutathione (10 to 15 mM). Na2S2O4, not known as an effective reducing agent of disulfides, prevented but was relatively ineffective in reversing activation after it had been induced by PGG2. Pretreatment of the enzyme preparation with increasing concentrations of N-ethylmaleimide in the range of 0.01 to 1.0 mM prevented activation by PGG2 without affecting basal guanylate cyclase activity. These observations indicate that fatty acid hydroperoxides and prostaglandin endoperoxides promote activation of the cyclase by oxidation of enzyme-related thiol functions. In contrast PGE2, PGF2a, hydroxy fatty acids (13-OH-18:2, 12-OH-20:4) as well as saturated (18:0) monoenoic (18:1), dienoic (18:2), and tetraenoic (20:4) fatty acids were ineffective in promoting cyclase activation in the range of 1 to 10 micrometer. Studies to identify the species of the rapidly metabolized prostaglandin endoperoxides that serve as effectors of the cyclase indicated that PGG2 but not 15-OOH-PGE2 (the major buffer-rearrangement product of PGG2) is most likely an activator. In the case of PGH2, a rapidly generated (30 s) metabolite of PGH2 was found which contained a hydroperoxy or endoperoxy functional group and was equally as effective as PGH2 as an apparent activator of the enzyme. The combined effects of PGG2 and dehydroascorbic acid, another class of activator, exhibited additivity with respect to the rate at which the time-dependent activation was induced. These results suggest that activation of soluble guanylate cyclase from splenic cells can be achieved by the oxidation of sulfhydryl groups that may be associated with specific hydrophobic sites of the enzyme or a related regulatory component.

Animals↗
PubMed1978

Prostaglandin endoperoxides promote calcium release from a platelet membrane fraction in vitro.

A calcium sequestering platelet membrane fraction was prepared and the effect of arachidonic acid, PGG2 and PGH2 on calcium content evaluated. At 4 degrees C, 6.7--16.7 micrometers arachidonic acid caused significant release of calcium from preloaded vesicles. Such release was completely inhibited by aspirin pretreating the platelets from which the membrane fraction was prepared. Gamma-linolenic acid, not a substrate for prostaglandin synthesis, did not cause calcium release. At 37 degrees C, after a 5 minute calcium loading of the membrane vesicles, arachidonic acid, PGG2, and PGH2 caused release of calcium. Calcium release by the PGG2 and PGH2 was only slightly inhibited by aspirin. Imidazole, which prevented conversion of the prostaglandin endoperoxides to thromboxanes, also only slightly inhibited calcium release. Other prostaglandins including PGD2, PGE1, PGE2 and PGD2 had no effect on the calcium content of the vesicles. These studies suggest that PGG2 and PGH2 may exert their effects on platelets by mobilizing calcium from an internal membrane store to make it available to promote platelet activation.

Arachidonic Acids↗
PubMed1976

A shift from phospholipid to triglyceride synthesis when cell division is inhibited by trans-fatty acids.

The yeast mutant Saccharomyces cerevisiae (KD 46) requires added unsaturated fatty acid for growth. When cell growth was inhibited by the presence of trans-acids there was a marked inhibition of oleate esterification into phospholipids accompanying continued incorporation into triglycerides. Apparently some control point in phospholipid synthesis associated with the cell cycle occurs after the stage of phosphatidate biosynthesis.

Cell Division↗