Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.31 (beta-glucuronidase)
 7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The optimal reaction conditions and kinetic properties of eleven leukocyte acid hydrolases determined with the use of fluorigenic derivatives of 4-methyl-umbelliferone are described. The enzymes studied were acid phosphatase, aryl sulfatase, alpha- and beta-glucosidase, alpha- and beta-galactosidase, alpha-mannosidase, N-acetyl-beta-glucosaminidase, N-acetyl-beta-galactosaminidase, beta-glucuronidase and alpha-fucosidase. More than 90% of the activity of each enzyme was released into a 27,000 X g supernatant by a double sonication procedure employing 0.9% sodium chloride and 0.1% Triton X-100. The Km values obtained were similar to those previously reported for chromogenic subtrates. A single Km value could not be derived for beta-galactosidase because its double reciprocal plot was not linear. All enzymes could be measured with less than 10 mug of protein within 15 min. Activators and inhibitors studied included the chloride salts of Na+, K+, Zn2+, Ca2+, Mg2+, Hg2+, and Fe2+ as well as p-chloromercuriphenysulfonate, glutathione, BAL, EDTA, EGTA, Triton X-100 and sodium taurocholate. The reaction conditions described in this report can be used for the diagnosis of various lysosomal storage diseases and should facilitate the development of automated procedures for the analysis of these eleven enzyme activities with small quantities of blood.
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PMID:Human leukocyte acid hydrolases: characterization of eleven lysosomal enzymes and study of reaction conditions for their automated analysis. 0 26

The discovery of two previously unknown progesterone conjugates in human urine is presented. Urine from near-term pregnant women was applied to an anion exchange column and then eluted with an increasing concentration gradient of sodium chloride. Aliquots of collected fractions were subjected to four different treatments: (1) immediate extraction, (2) incubation at 37 degrees C, (3) beta-glucuronidase hydrolysis and (4) acid hydrolysis. All eluent aliquots were then further purified on a thin layer chromatographic system and measured for progesterone content by radioimmunoassay. The results suggest that progesterone, a non-hydroxylated delta 4-3 ketosteroid, occurs in at least two negatively charged conjugated forms in human urine. Comparison with elution patterns of known testosterone conjugates shows that these conjugates of progesterone travel on an anion exchange column similarly to testosterone glucuronide and testosterone sulphate. Although the exact structure of these progesterone conjugates remains to be elucidated, the property of physiological conjugation of progesterone has not previously been described. Quantitatively each of these conjugates appears to exist in amounts equal to or greater than the levels of free (unconjugated) urinary progesterone.
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PMID:Progesterone is conjugated in human urine. 737 Nov 85

Seven methods of erythrocyte entrapment--six by hypotonic exchange loading and one by chlorpromazine-induced endocytosis--were evaluted for (1) efficiency of incorporation of beta-glucuronidase, inulin, and glucose; (2) presence of beta-glucuronidase on the erythrocyte surface, as detected by hemagglutination or complement-dependent lysis in the prsence of antibody to beta-glucuronidase; (3) in vitro leakage of entrapped markers; and (4) morphologic alterations by scanning electron microscopy. Dependent on the method of entrapment, the incorporation of markers ranged from 0.7% to 6.2% for beta-glucuronidase, 3.4% to 31% for inulin, and 1.2% to 12.2% for glucose. Maximal incorporation by hypotonic exchange loading occurred when the initial concentration of sodium chloride (150 mM) was reduced to 50 or 75 mM. However, beta-glucuronidase was detected on the erythrocyte surface by hemagglutination for these methods as well as a dialysis method of loading. Essentially no leakage of entrapped enzyme was detected (< 1%) for all methods, although up to 11% of entrapped glucose was released during a 3 hr incubation at 37 degrees C in buffered whole blood. Finally, entrapment methods requiring the greatest reduction in salt concentration resulted in the formation of echinocytes, whereas stomatocytes were observed after entrapment by methods requiring lesser salt dilutions. These results demonstrate the efficiency of entrapment and relative integrity of erythrocytes following various loadng procedures and suggest that in vitro assessment may provide a useful predictor of the imunogenicity and in vivo fate of erythrocyte-entrapped enzymes or other therapeutic agents.
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PMID:Enzyme therapy XIV. Comparison of methods for enzyme entrapment in human erythrocytes. 740 Jun 65

A method has been developed for the simultaneous determination of antipyrine and its three major metabolites in plasma of patients with renal failure. Plasma samples (500 microl) were hydrolyzed with beta-glucuronidase/aryl sulphatase. The compounds, after addition of sodium chloride, were extracted with chloroform:ethanol (90:10, v/v) in acidic medium. Chromatographic conditions comprise a C18 column, a mobile phase with 30% methanol and 70% 0.25N sodium acetate buffer (pH 5.0), a total run time of 10 minutes, and ultraviolet absorbance detection at 254 nm. Confidence limits showed 0.5 to 40.0 microg/ml(-1) linearity (r2 = 0.999); 0.1 microg/ml(-1) HMA, 0.05 microg/ml(-1) antipyrine and NORA, and 0.5 microg/ml(-1) OHA sensitivity and absolute recovery >95%. Interprecision and intraprecision expressed as coefficient of variation were <10% for all compounds investigated. The assay shows to be suitable for pharmacokinetics and drug metabolism studies after administration of a single oral dose of 500 mg of antipyrine to a patient with hypertension and chronic renal failure (CL(CR) = 34.17 ml/min(-1); 1.73 m(-2)).
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PMID:Determination of antipyrine and metabolites in plasma of a patient with mild renal failure. 942 Nov 15

While the beta-glucuronidase activity of intact cells of Clostridium perfringens was higher in 0.95% sodium chloride (NaCl) than that in 0, 0.1 or 0.5%, that of Escherichia coli was higher in 0.1% NaCl than that in 0, 0.5 or 0.95% NaCl in 0.1 mol l-1 KH2PO4. However, the enzyme activity of both species of intact cells was higher in buffer containing 16 mEq sodium, 134 mEq potassium and 16 mEq chloride per litre than in that containing 146 mEq sodium, 13 mEq potassium and 146 mEq chloride. These findings suggest that bacterial cells are affected by the presence of NaCl and that the effect of NaCl on the activity of bacterial beta-glucuronidase may differ by location in the large intestine.
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PMID:Influence of sodium chloride on the beta-glucuronidase activity of Clostridium perfringens and Escherichia coli. 1097 40

Homologous recombination (HR) as a strand break repair mechanism was shown to be influenced by various factors. The balance of different vitamins, macro- and microelements, light spectrum, sodium chloride concentration as well as various environmental mutagens were shown to influence the frequency of HR. In this paper we analysed the influence of temperature (4, 22, and 32 degrees C) and day/night duration on the genome stability of plants. We analyzed the HR frequency in transgenic Arabidopsis thaliana plants carrying beta-glucuronidase based homologous recombination substrate. To find the recombination rate (RR), we related the HR frequency to the number of genomes present in plants grown under different conditions. The RR was also standardized to the transcription activity of the transgene. We found RR to be higher in plants grown at suboptimal temperatures (either 4 or 32 degrees C) as compared to plants grown at 22 degrees C. This negatively correlated with the plant metabolic rate and positively correlated with concentration of peroxide produced by plant. In contrast, the RR in plants grown at different day length (8-24 h) was the lowest in plants grown at the longest day (24 h) and highest in the plants grown at the shortest day (8 h). Over 15-fold difference in the RR between plants grown at the shortest and the longest day was observed. Such a difference in recombination rate was primarily due to the higher transgene activity and higher endoreduplication levels in plants grown at longer days. Our data suggests that even "moderate" changes of environmental conditions may have a significant effect on plant genome stability.
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PMID:Homologous recombination in plants is temperature and day-length dependent. 1579 Apr 91

A 1,993 bp region upstream of the gene encoding the betaine aldehyde dehydrogenase (BADH) was isolated from Suaeda liaotungensis K., and the analysis of the promoter sequence has revealed the existence of several putative cis-elements by the PLACE database. In this study, according to the characteristic of the BADH promoter, five chimeric constructs varied in the length of promoter fragments from -1,993, -1,466, -1,084, -573 and -300 to +62 bp relative to the transcriptional start site were placed to the upstream of the beta-glucuronidase (GUS) coding region and transferred to Nicotiana tabacum L.cv.89 by Agrobacterium tumefaciens-mediated leaf-disc transformation. The functional properties of each promoter fragment were examined by GUS histochemical staining and fluorescence quantitative analyses in the transgenic tobacco leaves treated with different NaCl concentrations for 48 h. The results show that healthy transgenic plants had decreased GUS activity in leaves, whereas a higher GUS activity was observed when the transgenic plants were challenged with sodium chloride (NaCl). Induction levels were proportional to the concentration of NaCl treatment, allowing fine-tuning of protein expression. GUS enzyme activity was enhanced 6.3-fold in transgenic tobacco leaves containing -300 bp promoter fragment in the presence of 400 mmol/l NaCl compared to the noninductive leaves. This suggests that the smallest promoter fragment (-300 to +62 bp) possesses all the essential cis-acting elements and is sufficient for NaCl induction.
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PMID:Functional analysis of BADH gene promoter from Suaeda liaotungensis K. 1792 16

Dicistronic binary vector constructs based on pGreenII vectors for Agrobacterium mediated gene transfer alleviate the translational expression monitoring of a target gene in plants. The functionality of the transformation vectors was proven by marker gene constructs containing a mannopine synthase promoter (p-MAS) fused to a beta-glucuronidase (gus) gene followed by an internal ribosome entry site and a firefly luciferase (luc) gene. The cap-dependent translation of a physically independent target protein can be monitored by the cap-independently co-translated luciferase, because both mRNAs are located on the same strand. Among three different IRES elements, the tobamo IRES element showed highest activity in transient expression. As a proof of principle for physiological studies the gus gene was replaced by a sodium antiporter gene (Atnhx1). Comparative studies with Atnhx1 transgenic luc expressing tobacco cell cultures and pea plants (Pisum sativum L.) showed improved salt tolerance in relation to their wild type counterparts grown under corresponding conditions. A coincidence of the luc gene expression and increased sodium chloride tolerance is demonstrated by measurement of luminescence and cell growth.
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PMID:Dicistronic binary vector system-A versatile tool for gene expression studies in cell cultures and plants. 1983 18