Linked Life Data

11141059

Source:http://linkedlifedata.com/resource/pubmed/id/11141059

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2001-1-25
pubmed:abstractText
BphF is a small, soluble, Rieske-type ferredoxin involved in the microbial degradation of biphenyl. The rapid, anaerobic purification of a heterologously expressed, his-tagged BphF yielded 15 mg of highly homogeneous recombinant protein, rcBphF, per liter of cell culture. The reduction potential of rcBphF, determined using a highly oriented pyrolytic graphite (HOPG) electrode, was -157+/- 2 mV vs the standard hydrogen electrode (SHE) (20 mM MOPS, 80 mM KCl, and 1 mM dithiothreitol, pH 7.0, 22 degrees C). The electron paramagnetic resonance spectrum of the reduced rcBphF is typical of a Rieske cluster while the close similarity of the circular dichroic (CD) spectra of rcBphF and BedB, a homologous protein from the benzene dioxygenase system, indicates that the environment of the cluster is highly conserved in these two proteins. The reduction potential and CD spectra of rcBphF were relatively independent of pH between 5 and 10, indicating that the pK(a)s of the cluster's histidinyl ligands are not within this range. Gel filtration studies demonstrated that rcBphF readily oligomerizes in solution. Crystals of rcBphF were obtained using sodium formate or poly(ethylene glycol) (PEG) as the major precipitant. Analysis of the intermolecular contacts in the crystal revealed a head-to-tail interaction that occludes the cluster, but is very unlikely to be found in solution. Oligomerization of rcBphF in solution was reversed by the addition of dithiothreitol and is unrelated to the noncovalent crystallographic interactions. Moreover, the oligomerization state of rcBphF did not influence the latter's reduction potential. These results indicate that the 450 mV spread in reduction potential of Rieske clusters of dioxygenase-associated ferredoxins and mitochondrial bc(1) complexes is not due to significant differences in their solvent exposure.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
9
pubmed:volume
40
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
84-92
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11141059-Burkholderia, pubmed-meshheading:11141059-Circular Dichroism, pubmed-meshheading:11141059-Crystallography, X-Ray, pubmed-meshheading:11141059-Electrochemistry, pubmed-meshheading:11141059-Electron Spin Resonance Spectroscopy, pubmed-meshheading:11141059-Electron Transport, pubmed-meshheading:11141059-Electron Transport Complex III, pubmed-meshheading:11141059-Ferredoxins, pubmed-meshheading:11141059-Gene Expression Regulation, Bacterial, pubmed-meshheading:11141059-Genetic Vectors, pubmed-meshheading:11141059-Hydrolases, pubmed-meshheading:11141059-Iron-Sulfur Proteins, pubmed-meshheading:11141059-Oxidation-Reduction, pubmed-meshheading:11141059-Recombinant Proteins, pubmed-meshheading:11141059-Solutions, pubmed-meshheading:11141059-Spectrophotometry, Ultraviolet, pubmed-meshheading:11141059-Thermodynamics
pubmed:year
2001
pubmed:articleTitle
Characterization of BphF, a Rieske-type ferredoxin with a low reduction potential.
pubmed:affiliation
Department of Biochemistry, Université Laval, Québec, Canada.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't