Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.6.1 (sulfatase)
3,205 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two monoclonal antibodies (10C10 and 4D5) have been developed from the spleen cells of Balb/c mice immunized with 6-aminobenzo[a]pyrene covalently coupled to bovine serum albumin. These antibodies have been used in an immunoassay for the detection of benzo[a]pyrene and its metabolites in mouse urine. The antibodies were characterized in terms of sensitivity and specificity by competitive enzyme-linked immunosorbent assay (ELISA). With both antibodies, 50% inhibition of antibody binding is at 4 pmol of BP. The antibodies also cross-react with a number of BP metabolites as well as with several other polycyclic aromatic hydrocarbons (PAHs) including pyrene, 1-aminopyrene, and 7,12-dimethylbenz[a]anthracene but with different sensitivities. These results suggest that this assay will detect multiple PAH metabolites in urine. To test the assay on biological samples, mice were treated with [3H]BP, and urine was collected and digested with beta-glucuronidase and aryl sulfatase. Several methods were used to isolate BP and its metabolites from the urine, including ethyl acetate extraction, Sep-pak C18 cartridge chromatography, XAD2 resin chromatography, and immunoaffinity chromatography with antibody 4D5. Analysis of the urine extracts with antibody 4D5 gave 50% inhibition at 12-15 pmol of metabolites. Thus, quantitation of metabolites in this sample by competitive ELISA against a standard curve of BP would have underestimated actual metabolite levels by about 70%. This assay will be applied to the analysis of urines from individuals with environmental or occupational exposure. Since humans are usually exposed to BP in complex mixtures of PAHs, multiple metabolites may be present in the urine, making absolute quantitation difficult. This assay should thus serve as a general indicator of exposure to this class of chemicals.
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PMID:Immunologic methods for the detection of benzo[a]pyrene metabolites in urine. 213 77

Investigations were carried out on the intracellular fate of formaldehyde treated bovine serum albumin (F-BSA), in liver non-parenchymal cells. This paper reports the observations and results obtained by us. The first part of our work involved the injecting of the compound into either a) normal rats, b) rats injected with Triton WR 1339 or c) rats treated with mannan. Fractions obtained after differential and isopycnic centrifugation in sucrose gradients, were analysed by SDS-gel electrophoresis and fluorography. The degradation takes place in a two step process. The molecule is first split into radiolabeled compounds that are still acid precipitable. This is followed by the appearance of acid soluble radioactive molecules. In a sucrose gradient the first kind of degradation products exhibit a distribution totally different from that of acid soluble degradation compounds. In the second part of our experiments, fairly pure fractions of the organelles, known to be involved in the endocytic pathway i.e. endosomes, transfer lysosomes and accumulation lysosomes (marked by the presence of either Triton WR 1339 or mannan) were isolated and incubated with [125I]-F-BSA. These experiments revealed that endosomes, isolated by us, are incapable of degradation. Accumulation lysosomes arising exclusively from liver non-parenchymal cells (in which mannan had accumulated) though rich in certain hydrolases eg. arylsulfatase did not have an efficient proteolytic machinery. Our results, both from in vivo and in vitro studies, suggest that the first degradation step occurs in one type of structure (probably not endosomes), a sort of hybrid endosome-lysosome (as they are not affected by glycyl-1-phenyl-2-napthylamide) and the second step in a different type of lysosomes, what we have designated transfer lysosomes.
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PMID:Intracellular degradation by liver endothelial cells. 262 58

Arylsulfatase B, purified to homogeneity from human eosinophils, is a tetrameric enzyme whose activity varied in accordance with the state of association of its monomeric subunits. The rate of dissociation of oligomeric forms was slow relative to the rate of the enzymatic reaction so that the kinetic properties of the enzyme depended on the concentration of the enzyme before assay. For concentrated enzyme solutions (14 micrograms/ml), Lineweaver-Burk analysis demonstrated substrate inhibition at greater than or equal to 20 mM substrate and revealed two distinct regions of activity at low and intermediate substrate concentrations. The addition of bovine serum albumin (60 micrograms/ml) or sucrose (0.25 M), which prevent subunit dissociation, yielded a linear relationship on Lineweaver-Burk analysis at non-inhibitory substrate concentrations. For dilute enzyme concentrations (4.7 micrograms/ml), inhibition occurred at greater than or equal to 2 mM substrate. Nanomolar amounts of leukotriene C4 (LTC4), relative to millimolar concentrations of substrate, inhibited eosinophil arylsulfatase B. On Lineweaver-Burk analysis, the pattern of inhibition of LTC4 with concentrated enzyme was compatible with competitive inhibition of only one oligomeric form of the enzyme, whereas at low enzyme concentrations the pattern of inhibition was apparently competitive. These findings suggest that LTC4 is a potent competitive inhibitor of a dissociated, possibly dimeric, form of the enzyme. Nanomolar concentrations of LTC4, leukotriene D4, and leukotriene E4 were equally inhibitory, whereas leukotriene B4 and isomeric 5,12-dihydroxyeicosatetraenoic acids had no inhibitory activity, indicating a requirement for a thiopeptide at C-6. Thiopeptide leukotriene analogs without an intact triene structure also lacked inhibitory activity. Sulfoxide analogs of LTC4 and leukotriene D4 were potent inhibitors, although two sulfone analogs of leukotriene D4 were not inhibitory. Arylsulfatase B did not inactivate the spasmogenic activity of sulfidopeptide leukotrienes. These findings indicate that sulfidopeptide leukotrienes and their sulfoxide derivatives may regulate by competitive inhibition the activity of oligomeric forms of the eosinophil lysosomal hydrolase, arylsulfatase B.
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PMID:Inhibition of homogeneous human eosinophil arylsulfatase B by sulfidopeptide leukotrienes. 300 83

A sensitive and specific RIA has been developed to measure thyronine (To) in urine. The RIA used an anti-To antibody obtained from a rabbit immunized with a L-To-human serum albumin conjugate and [3H]To as the radioligand. The acetic acid analog of To (ToAc), that is the diphenyl structure with an acetic acid side-chain, cross-reacted strongly with the antibody. Relative to To, it cross-reacted 160% in phosphate-buffered saline, pH 7.4, and 100% in 0.075 mol/L barbital buffer, pH 8.6, containing sodium salicylate (final concentration, 8 mg/mL). The latter conditions were employed for the RIA, and the results reported thus reflect the presence of To and/or ToAc. 3-Monoiodothyronine, 3'-monoiodothyronine, 3',5'-diiodothyronine, and 3,5-diiodothyronine cross-reacted with the anti-To antibody 1.9%, 1.7%, 0.3%, and 0.2%, respectively; the cross-reactivity of other To derivatives and tyrosine and its derivatives was less than 0.05%. Urinary To and/or ToAc excretion in 12 normal subjects averaged 16 +/- 2 (+/- SE) micrograms/day (59 +/- 9 nmol/day) or 14 +/- 2 micrograms/g creatinine (5.9 +/- 0.6 nmol/mmol creatinine). Treatment of urine from normal subjects with beta-glucuronidase or sulfatase did not significantly alter the To content. Column and thin layer chromatographic studies revealed that 83% and 61%, respectively (range, 37-100%), of urinary To immunoreactivity was attributable to ToAc. The mean daily excretion of To in 20 patients with nonthyroidal illness [NTI; 22 +/- 4 micrograms/day (82 +/- 17 nmol/day)] was similar to that in normal subjects, but was elevated when expressed as nanomoles per mmol creatinine (20 +/- 2; P less than 0.001), because creatinine excretion was reduced in the NTI patients. The mean daily urinary To excretion in 13 patients with hyperthyroidism due to Graves' disease was slightly elevated [29 +/- 6 micrograms/day (108 +/- 21 nmol/day); P less than 0.1], but was clearly elevated when expressed as nanomoles per mmol creatinine (37 +/- 8; P less than 0.001), again because creatinine excretion was reduced in these patients. The mean urinary To excretion was subnormal in 13 patients with hypothyroidism and was significantly (P less than 0.005) less than that in the NTI patients regardless of the manner in which the results were expressed. Analysis of pronase hydrolysates of thyroid glands obtained at autopsy from euthyroid patients suggested that the To content of the thyroid approximates only 1.2% that of T4, supporting the thesis that prior iodination of tyrosine is critical for the coupling process in the thyroid.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:A radioimmunoassay for measurement of thyronine and its acetic acid analog in urine. 341 Sep 34

Thirty-three strains of Vibrio vulnificus of clinical and environmental origin were examined for production of 12 extracellular enzymes of potential importance to the virulence of this bacterium. Strains of Vibrio vulnificus were consistent in their production of protease, mucinase, lipase, chondroitinase, hyaluronidase, DNase, sulfatase, and hemolysin. No differences between clinical and environmental isolates were noted. Although none of the enzymes appeared to correlate with the ability of these strains to produce lethality in mice, the production of hemolysin and of a protease with activity against native serum albumin may be significant in the pathogenesis of the potentially fatal infections produced by this organism. The production of several of these exoenzymes also appeared to correlate with pathogenicity in the seven other Vibrio species examined. Culture filtrates of all virulent strains of Vibrio vulnificus were cytotoxic for Chinese hamster ovary cells, whereas those of the strains of Vibrio parahaemolyticus and Vibrio alginolyticus examined lacked this activity.
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PMID:Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus. 352 90

Estrone and dehydroepiandrosterone (DHEA) sulfatases were studied in livers of normal and cirrhotic men. Their Km were 3.2 microM and 1.2 microM respectively. The microsomal sulfatases were solubilized by Miranol H2M and ultrasound. After gel filtration, the soluble material gave a single peak of activity for both substrates with a molecular weight of approximately 330,000. In terms of pmol of product.min-1 per mg of fresh tissue, the mean (+/- SD) values of estrone and DHEA sulfatase activities were lower in cirrhotic livers [(n = 7) (4.09 +/- 2.90 and 0.38 +/- 0.20)] than in normal livers [(n = 13)(8.29 +/- 4.00 and 0.69 +/- 0.20)]. The differences were statistically significant : p less than 0.03 for estrone sulfatase and p less than 0.01 for DHEA sulfatase. In cirrhotic men, the mean level of plasma estrone is increased whereas that of estrone sulfate is decreased. The variations may be related to the decrease of serum albumin in cirrhotic subjects.
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PMID:Steroid sulfatase activities in normal and cirrhotic livers and plasma levels of estrone sulfate, estrone and estradiol-17 beta in men. 609 55

Bovine serum albumin (BSA) in complete Freund's adjuvant (CFA) was injected into the knee joints of previously BSA-sensitized guinea pigs and rabbits. The primary immune reaction to BSA prevented secondary immune response to Trichinella larvae infection. We were unable to produce either eosinophilia in the peripheral blood or antibodies against Trichinella antigen (shown by complement fixation test). The additional injection of mediators of the inflammatory reaction or their precursors, e.g., complement component C5 and arachidonic acid, caused different histological pictures. C5 produced a prolonged acute inflammatory phase with abundant neutrophils, whereas arachidonic acid did not significantly change the inflammatory response as compared to controls. The additional application of eosinophil-enriched preparation (EEP) caused a conspicuously reduced influx of monocytes/macrophages, a reduction of lymphocyte numbers, a prolonged influx of neutrophils, increased arylsulfatase activity, earlier reduction of the inflammatory process, and earlier onset of synoviocyte regeneration as compared to controls.
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PMID:Experimental monoarthritis. Modulatory effect of injected eosinophils on influx of various types of inflammatory cells. 649 Jan 38

A significant decrease in total carbohydrates and particularly in mannose, galactose and sialic acid has been observed in vitamin A-deficient rat liver lysosomal membrane. These alterations adversely affect the membrane permeability and structure-linked latency of the lysosomal enzymes. Significant reduction in the pH-dependent in vitro binding of the lysosomal arylsulfatase B to the highly purified membrane has been observed in vitamin A deficiency. This is attributed to the decrease in electro-negativity, mainly due to the observed reduction in negatively-charged sialic acid residues on the outer side of the membrane. Similar reduction in sialic acid content on the inner side of the membrane affects the microenvironment in the lysosomes. Intralysosomal pH, measured by computing the proteolytic activity of lysed lysosomes and of phagolysosomes, endocytosed with denatured 131I-labelled human serum albumin, is slightly consistently higher in vitamin A-deficient groups compared to that in control alone. This is reflected in the low rate of degradation of the entrapped proteins in vitamin A deficiency. The possible physiological significance of the observations is discussed with special reference to the loss of surface carbohydrates, particularly sialic acid, in vitamin A-deficient rats.
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PMID:Alterations in surface carbohydrates and in some functional properties of liver lysosomal membrane in vitamin A-deficient rat. 721 69

A benztropine RIA based on polyclonal antisera raised in New Zealand white rabbits has been developed. The drug-protein conjugates employed had a variety of moles of benztropine hemisuccinate coupled per mole of protein (bovine serum albumin or bovine thyroglobulin). Six antisera were developed and the one with the highest titer was further evaluated for its cross reactivity to N-desmethylbenztropine (4%) and the antipsychotic agents fluphenazine, flupenthixol, chlorpromazine, and haloperidol (all < 1%). The selected antiserum demonstrated sufficient sensitivity to measure benztropine from 0.156 to 100 ng/mL plasma in a 200-microL plasma sample, with a mean CV of < 6%. The RIA was applied to the analysis of steady-state plasma samples obtained from patients undergoing treatment with benztropine and plasma samples obtained from human volunteers and dogs orally dosed with the drug. Both the human and dog plasma samples, when analyzed after hydrolysis with beta-glucuronidase/sulfatase, demonstrated increments in benztropine concentrations, suggesting the drug may be undergoing biotransformation to phase II metabolite(s). In addition, when benztropine was selectively extracted from the unhydrolyzed plasma samples, there was a significant decrease in drug level, which further suggested that the antiserum cross reacted with phase II metabolite(s). The shape of the plasma concentration versus time profile obtained from the dog studies suggested that the drug might also undergo enterohepatic recycling.
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PMID:Development of a sensitive and specific radioimmunoassay for benztropine. 825 87

We have examined the effect of chloroquine on rat liver lysosomal enzyme distributions after isopycnic centrifugation in a sucrose gradient. Chloroquine injection causes the large majority of cathepsin C, acid phosphatase and N acetyl glucosaminidase to migrate towards lower density regions; on the other hand only about 50% of arylsulfatase and acid deoxyribonuclease are subjected to such a density shift. To specifically mark hepatocyte lysosomes and sinusoidal cell lysosomes, rats were injected with galactosylated bovine serum albumin (A) or mannosylated bovine serum albumin (B) labelled with 125I tyramine cellobiose; A is selectively endocytosed by hepatocytes, B by sinusoidal cells. The radioactivity distribution is affected by chloroquine in the same way as cathepsin C, after injection of A though it is not influenced by chloroquine after the injection of B. These results show that chloroquine does not modify the density of liver sinusoidal cell lysosomes when it decreases the density of hepatocyte lysosomes. Such a difference could result from the fact that sinusoidal cell lysosomes do not accumulate chloroquine to the same extent as hepatocyte lysosomes. Chloroquine treatment could be useful to distinguish between hepatocyte lysosomes and sinusoidal cell lysosomes.
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PMID:Chloroquine allows to distinguish between hepatocyte lysosomes and sinusoidal cell lysosomes. 843 31


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