Erythrosephosphate and 6-phosphogluconate, classical inhibitors of the conventional PGIs with K i values in the micromolar range e. It should be noted that S. The vials were then cooled on ice for 5 min, and the residual enzyme activity was tested and compared to the controls unheated samples. Open in a separate window. Two distinct heterodisulfide reductase-like enzymes in the sulfate-reducing archaeon Archaeoglobus profundus. The deduced activation energy of the latter was significant lower than the corresponding values of the other cPGIs in the present study.
AfcPGI obtained from 0. Structure of native phosphoglucose isomerase from rabbit: Parameter Property a of: Thus far, conclusive biochemical studies analyzing the role of metals have not been performed. Sequence comparison and phylogenetic affiliation. The measured concentration profile of sedimentation-diffusion equilibrium runs were best fit by using a molecular mass of 27 kDa, which is about the calculated mass of the AfcPGI
Glucosephosphate isomerase from the hyperthermophilic pvis Methanococcus jannaschii: We describe here the identification and functional characterization of cPGIs from the euryarchaeota Archaeoglobus fulgidus and Methanosarcina mazeias well as the bacteria Salmonella enterica serovar Typhimurium and Ensifer meliloti.
Multiple sequence alignment of amino acid sequences of the cPGI family. All metals were used in the form of chlorides. New Year Celebrations – The PGI superfamily is found in all three domains—archaea, bacteria, and fornat probably has been present in the last common ancestor as well.
The unique features of glycolytic pathways in Archaea. Spectra were recorded before and after each temperature tyesis. Recombinant proteins were purified as follows. The following controls were used.
Home About Courses Ph. Thus far this family comprises nine euryarchaeal and formta proteobacterial sequences. Temperature dependence and thermal stability. Moreover, the presence of a divalent metal, most likely iron, as deduced from the crystal structure of PfcPGI bound with 6-phosphogluconate is a prerequisite in the recently proposed hydride transfer mechanism of PfcPGI The following standard conditions were used: The region between the two cupin motifs, which varies from a fomat of 11 amino acids in some microbial enzymes to more than amino acids in certain eukaryotic transcription factors and dioxygenases 1718comprises 20 to 31 amino acids in cPGIs.
University of Hanover, Hanover, Germany. Programmes – January.
The cPGIs from the mesophiles M. Crystal structure of rabbit phosphoglucose isomerase complexed with 5-phospho-d-arabinonate identifies the role of Glu in catalysis.
Pgis thesis format outline
DNA sequencing with chain-terminating inhibitors. The genome of Methanosarcina mazei: The genome of M. Crystal structure of rabbit phosphoglucose isomerase, a glycolytic enzyme that moonlights as neuroleukin, autocrine motility factor, and differentiation mediator.
Determination of metal content.
Postgraduate Institute of Agricultur
The following primers were used to introduce unique restriction sites for NdeI and XhoI, respectively: An alignment of all known cPGIs is given in Fig. The metal content measured depended on both the source of the protein and the purification procedure used due to different contaminating metal contents of the buffer systems used. After the column was washed with 60 pgks of buffer B and ml of buffer C 50 mM piperazine [pH 5.
In most cases the corresponding metal-binding site of other cupin superfamily proteins is coordinated by three histidines and a glutamate as well Recently, a novel type of PGI has been identified and characterized from the hyperthermophilic euryarchaeon Pyrococcus furiosus 2252 and later from the closely related Thermococcus litoralis Acknowledgments We thank C. The given pH values correspond to the respective temperature used.
Heat-induced unfolding of the respective cPGIs was analyzed in temperature gradient experiments by using closed cuvettes 0. The protein samples were dialyzed against 10 mM sodium phosphate buffer pH 7. Their presence is presumably the result of a lateral gene transfer from a Thermococcus -like species after the lines of E.
The vials were then cooled on ice for 5 min, and the residual enzyme activity was tested and compared to the controls unheated samples.