@article {1586, title = {Identification of a glycolytic regulon in the archaea Pyrococcus and Thermococcus.}, journal = {FEMS Microbiology Letters}, volume = {260}, year = {2006}, month = {2006 Jul}, pages = {69-76}, abstract = {

The glycolytic pathway of the hyperthermophilic archaea that belong to the order Thermococcales (Pyrococcus, Thermococcus and Palaeococcus) differs significantly from the canonical Embden-Meyerhof pathway in bacteria and eukarya. This archaeal glycolysis variant consists of several novel enzymes, some of which catalyze unique conversions. Moreover, the enzymes appear not to be regulated allosterically, but rather at transcriptional level. To elucidate details of the gene expression control, the transcription initiation sites of the glycolytic genes in Pyrococcus furiosus have been mapped by primer extension analysis and the obtained promoter sequences have been compared with upstream regions of non-glycolytic genes. Apart from consensus sequences for the general transcription factors (TATA-box and BRE) this analysis revealed the presence of a potential transcription factor binding site (TATCAC-N(5)-GTGATA) in glycolytic and starch utilizing promoters of P. furiosus and several thermococcal species. The absence of this inverted repeat in Pyrococcus abyssi and Pyrococcus horikoshii probably reflects that their reduced catabolic capacity does not require this regulatory system. Moreover, this phyletic pattern revealed a TrmB-like regulator (PF0124 and TK1769) which may be involved in recognizing the repeat. This Thermococcales glycolytic regulon, with more than 20 genes, is the largest regulon that has yet been described for Archaea.

}, keywords = {Base Sequence, Binding Sites, Genetic, Glycolysis, Molecular Sequence Data, Promoter Regions, Pyrococcus, Regulon, Thermococcus, Transcription Initiation Site}, issn = {0378-1097}, doi = {10.1111/j.1574-6968.2006.00292.x}, author = {van de Werken, Harmen J G and Verhees, Corn{\'e} H and Akerboom, Jasper and de Vos, Willem M and van der Oost, John} } @article {1592, title = {Crystal structure of Pyrococcus furiosus phosphoglucose isomerase. Implications for substrate binding and catalysis.}, journal = {The Journal of Biological Chemistry}, volume = {278}, year = {2003}, month = {2003 Aug 29}, pages = {33290-7}, abstract = {

Phosphoglucose isomerase (PGI) catalyzes the reversible isomerization between d-fructose 6-phosphate and d-glucose 6-phosphate as part of the glycolytic pathway. PGI from the Archaea Pyrococcus furiosus (Pfu) was crystallized, and its structure was determined by x-ray diffraction to a 2-A resolution. Structural comparison of this archaeal PGI with the previously solved structures of bacterial and eukaryotic PGIs reveals a completely different structure. Each subunit of the homodimeric Pfu PGI consists of a cupin domain, for which the overall structure is similar to other cupin domain-containing proteins, and includes a conserved transition metal-binding site. Biochemical data on the recombinant enzyme suggests that Fe2+ is bound to Pfu PGI. However, as catalytic activity is not strongly influenced either by the replacement of Fe2+ by a range of transition metals or by the presence or absence of the bound metal ion, we suggest that the metal may not be directly involved in catalysis but rather may be implicated in substrate recognition.

}, keywords = {Amino Acid, Amino Acid Sequence, Binding Sites, Chemical, Crystallography, Dimerization, Fructosephosphates, Glucose-6-Phosphate, Glucose-6-Phosphate Isomerase, Ions, Iron, Metals, Models, Molecular, Molecular Sequence Data, Protein Binding, Protein Conformation, Protein Structure, Pyrococcus furiosus, Secondary, Selenomethionine, Sequence Homology, Substrate Specificity, Temperature, Tertiary, X-Ray, X-Ray Diffraction}, issn = {0021-9258}, doi = {10.1074/jbc.M305170200}, author = {Berrisford, John M and Akerboom, Jasper and Turnbull, Andrew P and de Geus, Daniel and Sedelnikova, Svetlana E and Staton, Ian and McLeod, Cameron W and Verhees, Corn{\'e} H and van der Oost, John and Rice, David W and Baker, Patrick J} } @article {1591, title = {Purification, crystallization and preliminary crystallographic analysis of phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus.}, journal = {Acta Crystallographica. Section D, Biological Crystallography}, volume = {59}, year = {2003}, month = {2003 Oct 19}, pages = {1822-3}, abstract = {

The glycolytic enzyme phosphoglucose isomerase catalyses the reversible isomerization of glucose 6-phosphate to fructose 6-phosphate. The phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus, which shows no sequence similarity to any known bacterial or eukaryotic phosphoglucose isomerase, has been cloned and overexpressed in Escherichia coli, purified and subsequently crystallized by the hanging-drop method of vapour diffusion using 1.6 M sodium citrate as the precipitant at pH 6.5. Multiple-wavelength anomalous dispersive X-ray data have been collected to a maximum resolution of 1.92 A on a single selenomethionine-incorporated crystal. This crystal belongs to space group C2, with approximate unit-cell parameters a = 84.7

}, keywords = {Crystallization, Crystallography, Escherichia coli, Glucose-6-Phosphate Isomerase, Protein Conformation, Pyrococcus furiosus, Recombinant Proteins, Selenomethionine, X-Ray}, issn = {0907-4449}, author = {Akerboom, Jasper and Turnbull, Andrew P and Hargreaves, David and Fisher, Martin and de Geus, Daniel and Sedelnikova, Svetlana E and Berrisford, John M and Baker, Patrick J and Verhees, Corn{\'e} H and van der Oost, John and Rice, David W} } @article {1593, title = {Molecular and biochemical characterization of a distinct type of fructose-1,6-bisphosphatase from Pyrococcus furiosus.}, journal = {Journal of Bacteriology}, volume = {184}, year = {2002}, month = {2002 Jun}, pages = {3401-5}, abstract = {

The Pyrococcus furiosus fbpA gene was cloned and expressed in Escherichia coli, and the fructose-1,6-bisphosphatase produced was subsequently purified and characterized. The dimeric enzyme showed a preference for fructose-1,6-bisphosphate, with a K(m) of 0.32 mM and a V(max) of 12.2 U/mg. The P. furiosus fructose-1,6-bisphosphatase was strongly inhibited by Li(+) (50\% inhibitory concentration, 1 mM). Based on the presence of conserved sequence motifs and the substrate specificity of the P. furiosus fructose-1,6-bisphosphatase, we propose that this enzyme belongs to a new family, class IV fructose-1,6-bisphosphatase.

}, keywords = {Amino Acid Sequence, Archaeal Proteins, Cloning, DNA, Fructose-Bisphosphatase, Molecular, Molecular Sequence Data, Pyrococcus furiosus, Sequence Analysis, Substrate Specificity, Temperature}, issn = {0021-9193}, author = {Verhees, Corn{\'e} H and Akerboom, Jasper and Schiltz, Emile and de Vos, Willem M and van der Oost, John} }