Novel Selenium-containing Human Single-chain Variable Fragment with Glutathione Peroxidase Activity from Computer-aided Molecular Design-中国期刊网

摘要 Inordertoenhancetheglutathioneperoxidase(GPX)catalyticactivityoftheselenium-containingsingle-chainvariablefragments(Se-scFv),anovelhumanscFvwasdesignedonthebasisofthestructureofhumanantibodyandoptimizedviabioinformaticsmethodssuchashomologoussequenceanalysis,three-dimensional(3D)modelbuilding,binding-siteanalysisanddocking.TheDNAsequenceofthenewhumanscFvwassynthesizedandclonedintotheexpressionvectorpET22b(+),thenthescFvproteinwasexpressedinsolubleforminEscherichiacoliBL21(DE3)andpurifiedbyNi2+-immobilizedmetalaffinitychromatography(IMAC).TheserineresidueofscFvintheactivesitewasconvertedintoselenocysteine(Sec)withthechemicalmodificationmethod,thus,thehumanSe-scFvwithGPXactivitywasobtained.TheGPXactivityoftheSe-scFvproteinwascharacterized.ComparedwithotherSe-scFv,thenewhumanSe-scFvshowedsimilarefficiencyforcatalyzingthereductionofhydrogenperoxidebyglutathione.ItexhibitedpHandtemperaturedependentcatalyticactivityandatypicalping-pongkineticmechanism.

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Novel Selenium-containing Human Single-chain Variable Fragment with Glutathione Peroxidase Activity from Computer-aided Molecular Design

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Novel Selenium-containing Human Single-chain Variable Fragment with Glutathione Peroxidase Activity from Computer-aided Molecular Design

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