Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.5.1.18 (glutathione S-transferase)
 22,582 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A 567bp DNA fragment was amplified from Schistosoma japonicum adult worm mRNA by RT-PCR. Sequence analysis revealed that this fragment contained S. Japonicum Chinese strain membrane-associated protein (Sj-22.6) gene. Then this gene was cloned into the expression vector pGEX-4T, and subsequently expressed in Escherichia coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was about 48 kD. The yield of expression was around 40 mg/L E.coli culture. The immunological test suggested that the recombinant protein had good antigenity which could make a good basis for the research of its immunological function in Schistosomiasis.
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PMID:Cloning of Schistosoma japonicum Chinese strain 22.6kD membrane-associated protein (Sj-22.6) gene and its overproduction on Escherichia coli. 1066 29

Interleukin-10 (IL-10) cytokine production was assessed using peripheral blood mononuclear cells (PBMC) from 67 individuals living in an area endemic for schistosomiasis japonica in China (Dongting Lake, Hunan Province), and 11 control subjects from a non-endemic part of the same Province. Production of IL-10 was measured following in vitro stimulation of PBMC using whole parasite extract (SWAP) or a panel of recombinant Schistosoma japonicum antigens (22-kDa tegumental membrane-associated antigen, glyceraldehyde-3-phosphate dehydrogenase, paramyosin, 14-kDa fatty acid-binding protein and 28-kDa glutathione S-transferase) which are of recognized interest in the development of protective immunity to schistosomiasis. Significantly, PBMC isolated from the exposed population compared with the non-exposed population produced higher levels of IL-10. There was a trend towards higher mean levels of IL-10 release in putatively resistant (insusceptible) (consistently egg negative but highly exposed) individuals compared with susceptible (egg-positive) subjects from the exposed population. Analysis of individual exposure (the duration of water contact and the percent body surface area in contact with water, expressed as m2 h/day) vs. IL-10 production indicated a weak but consistent and statistically significant inverse correlation, with lower levels of exposure being associated with higher levels of IL-10. These results suggest an association between IL-10 production and resistance to S. japonicum in subjects from this Chinese population exposed to infection.
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PMID:Production of interleukin-10 by peripheral blood mononuclear cells from residents of a marshland area in China endemic for Schistosoma japonicum. 1126 78

DNA sequence coding for a portion of DNA binding protein (amino acids 3-58) of bovine adenovirus type-3 (BAV-3) was cloned and expressed in Escherichia coli as a fusion protein with Schistosoma japonicum glutathione S-transferase. The fusion protein was affinity purified and used to immunize rabbits. Immunoprecipitation and Western blot analysis showed that the antiserum could specifically recognize a protein of 48 kDa in BAV-3-infected cells, which was produced both in early and late phases of BAV-3 life cycle. Based on the ability of antiserum to recognize DNA binding protein, a novel assay for BAV-3 quantitation was established. The assay is less time consuming and can be performed on a wide variety of bovine cells. In addition, virus titers determined by this assay are comparable to the standard plaque assay.
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PMID:Determination of bovine adenovirus-3 titer based on immunohistochemical detection of DNA binding protein in infected cells. 1133 49

The binding of three competitive glutathione analogue inhibitors (S-alkylglutathione derivatives) to glutathione S-transferase from Schistosoma japonicum, SjGST, has been investigated by isothermal titration microcalorimetry at pH 6.5 over a temperature range of 15--30 degrees C. Calorimetric measurements in various buffer systems with different ionization heats suggest that no protons are exchanged during the binding of S-alkylglutathione derivatives. Thus, at pH 6.5, the protons released during the binding of substrate may be from its thiol group. Calorimetric analyses show that S-methyl-, S-butyl-, and S-octylglutathione bind to two equal and independent sites in the dimer of SjGST. The affinity of these inhibitors to SjGST is greater as the number of methylene groups in the hydrocarbon side chain increases. In all cases studied, Delta G(0) remains invariant as a function of temperature, while Delta H(b) and Delta S(0) both decrease as the temperature increases. The binding of three S-alkylglutathione derivatives to the enzyme is enthalpically favourable at all temperatures studied. The temperature dependence of the enthalpy change yields negative heat capacity changes, which become less negative as the length of the side chain increases.
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PMID:A calorimetric study of the binding of S-alkylglutathiones to glutathione S-transferase. 1145 43

The binding properties of a glutathione S-transferase (EC 2.5.1.18) from Schistosoma japonicum to substrate glutathione (GSH) has been investigated by intrinsic fluorescence and isothermal titration calorimetry (ITC) at pH 6.5 over a temperature range of 15-30 degrees C. Calorimetric measurements in various buffer systems with different ionization heats suggest that protons are released during the binding of GSH at pH 6.5. We have also studied the effect of pH on the thermodynamics of GSH-GST interaction. The behaviour shown at different pHs indicates that at least three groups must participate in the exchange of protons. Fluorimetric and calorimetric measurements indicate that GSH binds to two sites in the dimer of 26-kDa glutathione S-transferase from Schistosoma japonicum (SjGST). On the other hand, noncooperativity for substrate binding to SjGST was detected over a temperature range of 15-30 degrees C. Among thermodynamic parameters, whereas DeltaG degrees remains practically invariant as a function of temperature, DeltaH and DeltaS degrees both decrease with an increase in temperature. While the binding is enthalpically favorable at all temperatures studied, at temperatures below 25 degrees C, DeltaG degrees is also favoured by entropic contributions. As the temperature increases, the entropic contributions progressively decrease, attaining a value of zero at 24.3 degrees C, and then becoming unfavorable. During this transition, the enthalpic contributions become progressively favorable, resulting in an enthalpy-entropy compensation. The temperature dependence of the enthalpy change yields the heat capacity change (DeltaCp degrees ) of -0.238 +/- 0.04 kcal per K per mol of GSH bound.
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PMID:Thermodynamic analysis of the binding of glutathione to glutathione S-transferase over a range of temperatures. 1148 26

A cDNA fragment locating at the putative envelop protein 2(E2) region of GBV-C/HGV fused with Schistosoma japonicum, glutathione S-transferase(GST) was amplified with PCR from plasmid pGEX-E2. The amplified DNA fragment was inserted into plasmid pGEX-5X-1, at the downstream of the coding sequences of GST, in the same reading frame with the gene of GST. The fusion gene fragment of GST-E2 was amplified with PCR, using the recombinant plasmid pGEX-5X-1-E2 as the template. The amplified 1324 bp DNA fragment of GST-E2 was inserted into Pichia pastoris expression vector pPIC9K in reading frame with alpha-factor secreting signal peptide. The plasmid pPIC9K-GST-E2 was transformed into Pichia pastoris GS115 with electroporation. The transformants (His+ Muts) were selected and induced to express the 54kD GST-E2 fusion protein, which could be specially recognized by both the antisera directed against E2 and against GST. The GST-E2 fusion protein was purified with Sepharose 4B glutathione affinity chromatography to a purity of 95%. The expression was optimized to achieve the highest expression level of GST-E2 fusion protein which was accumulated up to 50% of total proteins in the culture supernatant. The GST-E2 protein derived from the recombinant Pichia pastoris was proved possessing antigenicity and high specificity by ELISA, probed with sera from the patients infected by GBV-C/HGV.
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PMID:[Expression and characterization of envelope protein 2 gene of hepatitis G virus in Pichia pastoris]. 1214 81

Triose-phosphate isomerase is an important candidate for schistosoma antigens. An 800 bp DNA fragment was amplified by RT-PCR from adult Schistosoma japonicum mRNA. Sequence analysis revealed that this fragment contained S. japonicum (Chinese strain) triose-phosphate isomerase gene. Then this gene was cloned into the expression vector pGEX-4T and subsequently expressed in E. coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was determined to be 54 kD. The yield of expression was around 30 mg/L E. coli culture. Western blotting showed that the recombinant protein had good antigenicity which could be helpful for the making of anti-S. japonicum multi-valent recombinant vaccine.
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PMID:Cloning of Schistosoma japonicum Chinese Strain TPI Gene and Characterization of Its Expression Product in Escherichia coli. 1216 13

A 600 bp DNA fragment was amplified by PCR, from an adult Schistosoma japonicum cDNA library. Sequence analysis revealed that this fragment containedthe S. japonicum Chinese Mainland strain fatty acid binding protein (Sj-14FABPc) gene. This gene was then cloned into the expression vector pGEX-2T, and subsequently expressed in Escherichia coli. The recombinant GST-fusion protein could be purified by glutathione agarose affinity chromatography. Its molecular weight was about 41 kD. The yield of expression was around 25 mg/L E. coli culture. The immunological test suggested that the recombinant protein had good antigenicity, and could be developed into a new vaccine molecule of S. japonicum.
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PMID:Cloning of Schistosoma japonicum Chinese Strain Fatty Acid Binding Protein (Sj-FABPc) Gene and Its Overproduction in Escherichia coli. 1217 84

The gene of the 28 kD glutathione S-transferase (GST) from the Chinese strain of Schistosoma japonicum had been expressed in the silkworm (Bombyx mori) cells and larvae by Bombyx mori nuclear polyhedrosis virus (BmNPV) vector which had been modified. The GST gene was inserted into the right position of the BmNPV genome as identified by Southern hybridization. The product was a 28 kD protein, had GST activity and antigenicity. The yield in BmN cells (1x10(6) cells/ml) was 0.77 mg and more than 5 mg in a silkworm larvae. All the results showed that it is possible to develop the GST expressed in the silkworm larvae to a new kind of genetic engineered schistosomiasis vaccine.
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PMID:Studies on the Expression of the 28 kD Glutathione S-transferase Gene from Schistosoma japonicum in the Silkworm (Bombyx mori) Larvae and Insect Cells. 1221 34

There has been some speculation about the salt independence of Schistosoma japonicum glutathione S-transferase (Sj26GST, EC. 2.5.1.18), but this aspect has not been carefully studied before. To establish the basis for a further development of this dependence, we have performed a methodical study of the influence of some important ions and their concentration on the binding properties of glutathione to Sj26GST by means of isothermal calorimetry and fluorescence quenching. Salts like NaCl, Na(2)SO(4) and MgSO(4) do not change practically the affinity of the protein for its substrate, whilst MgCl(2) has the effect of decreasing the affinity as its concentration rises. However, the enthalpy change is not affected by all the salts studied, and so, the entropy change is the causal factor in dropping the affinity. We also looked at the conformational stability of the protein under different conditions to check the structural changes they provide, and found that the unfolding parameters are practically not affected by the salt concentration. We discuss the results in terms of the chaotropic nature of the ions implied.
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PMID:Salt influence on glutathione--Schistosoma japonicum glutathione S-transferase binding. 1256 23


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