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dc.contributor.authorHofmann, Julia
dc.contributor.authorHeider, Christine
dc.contributor.authorLi, Wei
dc.contributor.authorKrausze, Joern
dc.contributor.authorRoessle, Manfred
dc.contributor.authorWilharm, Gottfried
dc.date.accessioned2013-10-31T12:11:40Z
dc.date.available2013-10-31T12:11:40Z
dc.date.issued2013-04
dc.identifier.citationRecombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF. 2013, 88 (2):243-7 Protein Expr. Purif.en
dc.identifier.issn1096-0279
dc.identifier.pmid23384479
dc.identifier.doi10.1016/j.pep.2013.01.010
dc.identifier.urihttp://hdl.handle.net/10033/304812
dc.description.abstractThe glycolytic enzyme pyruvate kinase (PK) generates ATP from ADP through substrate-level phosphorylation powered by the conversion of phosphoenolpyruvate to pyruvate. In contrast to other bacteria, Enterobacteriaceae, such as pathogenic yersiniae, harbour two pyruvate kinases encoded by pykA and pykF. The individual roles of these isoenzymes are poorly understood. In an attempt to make the Yersinia enterocolitica pyruvate kinases PykA and PykF amenable to structural and functional characterisation, we produced them untagged in Escherichia coli and purified them to near homogeneity through a combination of ion exchange and size exclusion chromatography, yielding more than 180 mg per litre of batch culture. The solution structure of PykA and PykF was analysed through small angle X-ray scattering which revealed the formation of PykA and PykF tetramers and confirmed the binding of the allosteric effector fructose-1,6-bisphosphate (FBP) to PykF but not to PykA.
dc.language.isoenen
dc.rightsArchived with thanks to Protein expression and purificationen
dc.subject.meshChromatography, Gelen
dc.subject.meshChromatography, Ion Exchangeen
dc.subject.meshEscherichia colien
dc.subject.meshGene Expressionen
dc.subject.meshGenetic Vectorsen
dc.subject.meshIsoenzymesen
dc.subject.meshModels, Molecularen
dc.subject.meshProtein Multimerizationen
dc.subject.meshPyruvate Kinaseen
dc.subject.meshRecombinant Proteinsen
dc.subject.meshScattering, Small Angleen
dc.subject.meshX-Ray Diffractionen
dc.subject.meshYersinia enterocoliticaen
dc.titleRecombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF.en
dc.typeArticleen
dc.contributor.departmentRobert Koch-Institute, Wernigerode Branch, Burgstr. 37, D-38855 Wernigerode, Germany.en
dc.identifier.journalProtein expression and purificationen
refterms.dateFOA2018-06-12T21:24:57Z
html.description.abstractThe glycolytic enzyme pyruvate kinase (PK) generates ATP from ADP through substrate-level phosphorylation powered by the conversion of phosphoenolpyruvate to pyruvate. In contrast to other bacteria, Enterobacteriaceae, such as pathogenic yersiniae, harbour two pyruvate kinases encoded by pykA and pykF. The individual roles of these isoenzymes are poorly understood. In an attempt to make the Yersinia enterocolitica pyruvate kinases PykA and PykF amenable to structural and functional characterisation, we produced them untagged in Escherichia coli and purified them to near homogeneity through a combination of ion exchange and size exclusion chromatography, yielding more than 180 mg per litre of batch culture. The solution structure of PykA and PykF was analysed through small angle X-ray scattering which revealed the formation of PykA and PykF tetramers and confirmed the binding of the allosteric effector fructose-1,6-bisphosphate (FBP) to PykF but not to PykA.


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