Citrate synthase from the hyperthermophilic Archaeon, Pyrococcus furiosus.
The gene encoding the enzyme citrate synthase has been cloned and sequenced from the hyperthermophilic Archaeon Pyrococcus furiosus, and the derived amino acid sequence has been phylogenetically compared with citrate synthases from archaeal, bacterial and eukaryal organisms. The gene has been over-expressed in Escherichia coli to produce an active enzyme that has then been characterized with respect to its kinetic, oligomeric and hyperthermostable properties. A structurally-based sequence alignment was made to the citrate synthase from the thermophilic Archaeon Thermoplasma acidophilum, the crystal structure of which we have determined recently. From this alignment, a homology-modelled structure for the P.furiosus citrate synthase was generated and analysed.
Muir
J M
JM
School of Biology and Biochemistry, University of Bath, UK.
Russell
R J
RJ
Hough
D W
DW
Danson
M J
MJ
D000595:Amino Acid Sequence; D001105:Archaea / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002950:Citrate (si)-Synthase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006358:Hot Temperature; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D002950:Citrate (si)-Synthase