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2013-02-04Zeitschriftenartikel DOI: 10.1016/j.pep.2013.01.010
Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF
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.accessioned2018-05-07T17:28:56Z
dc.date.available2018-05-07T17:28:56Z
dc.date.created2014-02-06
dc.date.issued2013-02-04none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reMstu0NfQV4Q/PDF/20jW5yErTe8E6.pdf
dc.identifier.urihttp://edoc.rki.de/176904/1816
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.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectEscherichia coli/geneticseng
dc.subjectGenetic Vectors/geneticseng
dc.subjectChromatography Geleng
dc.subjectChromatography Ion Exchangeeng
dc.subjectGene Expressioneng
dc.subjectIsoenzymes/chemistryeng
dc.subjectIsoenzymes/geneticseng
dc.subjectIsoenzymes/isolation & purificationeng
dc.subjectIsoenzymes/metabolismeng
dc.subjectModels Moleculareng
dc.subjectProtein Multimerizationeng
dc.subjectPyruvate Kinase/chemistryeng
dc.subjectPyruvate Kinase/geneticseng
dc.subjectPyruvate Kinase/isolation & purificationeng
dc.subjectPyruvate Kinase/metabolismeng
dc.subjectRecombinant Proteins/chemistryeng
dc.subjectRecombinant Proteins/geneticseng
dc.subjectRecombinant Proteins/isolation & purificationeng
dc.subjectRecombinant Proteins/metabolismeng
dc.subjectScattering Small Angleeng
dc.subjectX-Ray Diffractioneng
dc.subjectYersinia enterocolitica/chemistryeng
dc.subjectYersinia enterocolitica/enzymologyeng
dc.subjectYersinia enterocolitica/geneticseng
dc.subject.ddc610 Medizin
dc.titleRecombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10034891
dc.identifier.doi10.1016/j.pep.2013.01.010
dc.identifier.doihttp://dx.doi.org/10.25646/1741
local.edoc.container-titleProtein Expression and Purification
local.edoc.container-textHofmann, J., Heider, C., Li, W., Krausze, J., Roessle, M., Wilharm, G. Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF (2013) Protein Expression and Purification, 88 (2), pp. 243-247.
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.sciencedirect.com/science/article/pii/S1046592813000119
local.edoc.container-publisher-nameElsevier
local.edoc.container-volume88
local.edoc.container-issue2
local.edoc.container-year2013

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