| pubmed-article:1577013 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C0027021 | lld:lifeskim |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C0038774 | lld:lifeskim |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C0243071 | lld:lifeskim |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C1998793 | lld:lifeskim |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C0871161 | lld:lifeskim |
| pubmed-article:1577013 | lifeskim:mentions | umls-concept:C0591833 | lld:lifeskim |
| pubmed-article:1577013 | pubmed:issue | 3 | lld:pubmed |
| pubmed-article:1577013 | pubmed:dateCreated | 1992-6-9 | lld:pubmed |
| pubmed-article:1577013 | pubmed:abstractText | We previously constructed plasmids for synthesis of glutathione-peroxidase (GPx) mutants in an Escherichia coli expression system. In these recombinant proteins either cysteine ([Cys]GPx mutant) or serine ([Ser]GPx mutant) were present in place of the active-site selenocysteine (SeCys) of the natural enzyme. We have now investigated GPx activity of [Cys]GPx and [Ser]GPx mutants. Enzyme assays performed on preparations of these partially purified proteins demonstrated that the [Cys]GPx mutant exhibited a significant GPx activity, unlike the [Ser]GPx mutant. Purification of [Cys]GPx was performed in two steps of ion-exchange chromatography giving a 98% homogenous protein in 50% yield. The purified [Cys]GPx protein was shown to be a symmetrical tetramer by the means of gel-filtration HPLC and SDS/PAGE. Two isoelectric points were found (6.8 and 7.2) which may reflect two different oxidation states of the mutant protein. The GPx activity of the [Cys]GPx mutant was optimal at pH 8.5. The [Cys]GPx mutant had a specific activity approximately 1000-fold smaller than that of the natural enzyme, and was very easily inactivated by hydroperoxides. Inhibition of the activity with iodoacetate determined a pKa of 8.3, presumably that of the active-site cysteine. Unlike that of SeGPx, the GPx activity of [Cys]GPx was only slightly inhibited by mercaptosuccinate. We discuss hypothetical mechanistic constraints of either catalytic cycle, which may explain such results. | lld:pubmed |
| pubmed-article:1577013 | pubmed:language | eng | lld:pubmed |
| pubmed-article:1577013 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:1577013 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1577013 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:1577013 | pubmed:month | May | lld:pubmed |
| pubmed-article:1577013 | pubmed:issn | 0014-2956 | lld:pubmed |
| pubmed-article:1577013 | pubmed:author | pubmed-author:RocherCC | lld:pubmed |
| pubmed-article:1577013 | pubmed:author | pubmed-author:LalanneJ LJL | lld:pubmed |
| pubmed-article:1577013 | pubmed:author | pubmed-author:ChaudièreJJ | lld:pubmed |
| pubmed-article:1577013 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:1577013 | pubmed:day | 1 | lld:pubmed |
| pubmed-article:1577013 | pubmed:volume | 205 | lld:pubmed |
| pubmed-article:1577013 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:1577013 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:1577013 | pubmed:pagination | 955-60 | lld:pubmed |
| pubmed-article:1577013 | pubmed:dateRevised | 2007-7-23 | lld:pubmed |
| pubmed-article:1577013 | pubmed:meshHeading | pubmed-meshheading:1577013-... | lld:pubmed |
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| pubmed-article:1577013 | pubmed:meshHeading | pubmed-meshheading:1577013-... | lld:pubmed |
| pubmed-article:1577013 | pubmed:meshHeading | pubmed-meshheading:1577013-... | lld:pubmed |
| pubmed-article:1577013 | pubmed:meshHeading | pubmed-meshheading:1577013-... | lld:pubmed |
| pubmed-article:1577013 | pubmed:meshHeading | pubmed-meshheading:1577013-... | lld:pubmed |
| pubmed-article:1577013 | pubmed:year | 1992 | lld:pubmed |
| pubmed-article:1577013 | pubmed:articleTitle | Purification and properties of a recombinant sulfur analog of murine selenium-glutathione peroxidase. | lld:pubmed |
| pubmed-article:1577013 | pubmed:affiliation | Centre de Recherches Roussel-UCLAF, Romainville, France. | lld:pubmed |
| pubmed-article:1577013 | pubmed:publicationType | Journal Article | lld:pubmed |
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