2.50
Hdl Handle:
http://hdl.handle.net/10033/304812
Title:
Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF.
Authors:
Hofmann, Julia; Heider, Christine; Li, Wei; Krausze, Joern; Roessle, Manfred; Wilharm, Gottfried
Abstract:
The 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.
Affiliation:
Robert Koch-Institute, Wernigerode Branch, Burgstr. 37, D-38855 Wernigerode, Germany.
Citation:
Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF. 2013, 88 (2):243-7 Protein Expr. Purif.
Journal:
Protein expression and purification
Issue Date:
Apr-2013
URI:
http://hdl.handle.net/10033/304812
DOI:
10.1016/j.pep.2013.01.010
PubMed ID:
23384479
Type:
Article
Language:
en
ISSN:
1096-0279
Appears in Collections:
Publications from Division of Molekulare Struktur Biologie (MOSB)

Full metadata record

DC FieldValue Language
dc.contributor.authorHofmann, Juliaen
dc.contributor.authorHeider, Christineen
dc.contributor.authorLi, Weien
dc.contributor.authorKrausze, Joernen
dc.contributor.authorRoessle, Manfreden
dc.contributor.authorWilharm, Gottfrieden
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.en
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

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