1. osnaDocs
  2. Fachbereich 05 - Biologie / Chemie
  3. FB05 - E-Dissertationen

Characterization of the KdpFABC complex from Escherichia coli, of soluble subdomains from KdpB, and of a homologous protein of Methanococcus jannaschii

Bitte benutzen Sie diese Kennung, um auf die Ressource zu verweisen:
https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2003071014
Open Access logo originally created by the Public Library of Science (PLoS)
Langanzeige der Metadaten
DC ElementWertSprache
dc.creatorBramkamp, Marc
dc.date.accessioned2010-01-30T14:39:57Z
dc.date.available2010-01-30T14:39:57Z
dc.date.issued2003-07-10T15:32:58Z
dc.date.submitted2003-07-10T15:32:58Z
dc.identifier.urihttps://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2003071014-
dc.description.abstractThe KdpFABC complex is a P-type ATPase. Several features make the inducible Kplus-transporting ATPase a unique member of this enzyme family. Aspects of structure and function of KdpB, the catalytic subunit of the complex, were examined here. Site-directed mutagenesis of the charged residues aspartate 583 and lysine 586 in the putative transmembrane helix 5 of KdpB revealed that these charges are involved in the coupling of ATP hydrolysis to ion translocation. The binding of FITC was shown to be specific and the binding site is within the nucleotide-binding domain of KdpB, most probably at lysine 395. Modification of KdpB with FITC was affected by adenosine nucleotides. A Mg2plus-dependent hydrolysis of p-nitrophenolphosphate was observed, which was inhibited by micromolar concentrations of ortho-vanadate and FITC. Low concentrations of ATP stimulated pNPP hydrolysis, while higher concentrations of ATP were inhibitory. ADP, AMP and Pi inhibited the pNPP hydrolysis. The catalytic modules of KdpB were separately synthesized, purified and biochemically characterized. It was found that KdpBN was highly soluble and could be concentrated up to 1 mM and higher. Therefore, the KdpBN domain was used for further structural analysis using nuclear magnetic resonance spectroscopy. The KdpBN domain could be purified from cells grown in minimal medium with 15N-ammonium sulfate and 13C1-6 glucose as sole nitrogen and carbon sources, respectively. The purified, labeled KdpBN protein was applied to NMR analysis. High quality multidimensional NMR spectra were obtained (M. Haupt and H. Kessler, TU Munich, personal communication) and structure calculations leading to backbone assignments were carried out. An ortholog of the H4H5 domain of KdpB from the thermopilic archaeon Methanococcus jannaschii, Mj0968, was cloned, expressed, purified, and characterized.eng
dc.language.isoeng
dc.subjectKdp
dc.subjectpotassium transport
dc.subjectNMR
dc.subjectprotein structure
dc.subject.ddc570 - Biowissenschaften, Biologieger
dc.titleCharacterization of the KdpFABC complex from Escherichia coli, of soluble subdomains from KdpB, and of a homologous protein of Methanococcus jannaschiieng
dc.typeDissertation oder Habilitation [doctoralThesis]-
thesis.locationOsnabrück-
thesis.institutionUniversität-
thesis.typeDissertation [thesis.doctoral]-
thesis.date2003-02-11T12:00:00Z-
elib.elibid226-
elib.marc.edtfangmeier-
elib.dct.accessRightsa-
elib.dct.created2003-06-19T10:55:22Z-
elib.dct.modified2003-07-10T15:32:58Z-
dc.contributor.refereeProf. Dr. K. Altendorf
dc.contributor.refereeProf. Dr. W. Schoner
dc.subject.dnb30 - Chemieger
dc.subject.dnb32 - Biologieger
vCard.ORGFB5ger
Enthalten in den Sammlungen:FB05 - E-Dissertationen

Dateien zu dieser Ressource:
Datei Beschreibung GrößeFormat 
E-Diss226_thesis.pdfPräsentationsformat7,43 MBAdobe PDF
E-Diss226_thesis.pdf
Miniaturbild
Öffnen/Anzeigen
Zur Kurzanzeige


Alle Ressourcen im Repositorium osnaDocs sind urheberrechtlich geschützt, soweit nicht anderweitig angezeigt. rightsstatements.org