2013-02-04Zeitschriftenartikel DOI: 10.1016/j.pep.2013.01.010
Recombinant production of Yersinia enterocolitica pyruvate kinase isoenzymes PykA and PykF
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.
No license information
Show related Items with similar Title, Author, Creator or Subject.
2013-08-08ZeitschriftenartikelCytomegalovirus Downregulates IRE1 to Repress the Unfolded Protein Response Stahl, Sebastian; Burkhart, Julia M.; Hinte, Florian; Tirosh, Boaz; Mohr, Hermine; Zahedi, René P.; Sickmann, Albert; Ruzsics, Zsolt; Budt, Matthias; Brune, WolframDuring viral infection, a massive demand for viral glycoproteins can overwhelm the capacity of the protein folding and quality control machinery, leading to an accumulation of unfolded proteins in the endoplasmic reticulum ...
2005-06-20ZeitschriftenartikelViral promoters can initiate expression of toxin genes introduced into Escherichia coli Lewin, Astrid; Mayer, Martin; Chusainow, Janet; Jacob, Daniela; Appel, BerndBackground: The expression of recombinant proteins in eukaryotic cells requires the fusion of the coding region to a promoter functional in the eukaryotic cell line. Viral promoters are very often used for this purpose. ...
2013-04-15ZeitschriftenartikelGiardia duodenalis arginine deiminase modulates the phenotype and cytokine secretion of human dendritic cells by depletion of arginine and formation of ammonia Banik, Stefanie; Viveros, Pablo Renner; Seeber, Frank; Klotz, Christian; Ignatius, Ralf; Aebischer, ToniDepletion of arginine is a recognized strategy that pathogens use to evade immune effector mechanisms. Depletion depends on microbial enzymes such as arginases, which are considered virulence factors. The effect is mostly ...