Author information Article notes Copyright and License information Disclaimer. Identification, cloning and functional overexpression of cPGIs. Accession numbers were as follows: To prove their coding function as cpgi , these ORFs were cloned and functionally overexpressed in Escherichia coli as follows. In sedimentation velocity experiments the apparent sedimentation coefficient s app 20,W and an approximated molecular mass were evaluated from the velocity and shape of the sedimenting boundary by fitting the time-dependent concentration profiles calculated with Lamm’s differential equation for a single sedimenting species to the measured data by using the program package AKKUPROG Phylogenetic analyses suggest that bacterial cPGIs originated from euryarchaeota by lateral gene transfer.
Low buffer concentrations 10 mM and Ultrapure Milli-Q water had to be used for all substances in the assays. The thermophilic properties of the cPGIs were analyzed by determining the temperature dependence of the specific activity by time-dependent heat inactivation of the proteins Fig. Two consensus sequences for the novel cPGI family were derived. I really valued the support and encouragement received from lecturers and my colleagues. Received Jul 29; Accepted Sep
Structure of oxalate decarboxylase from Bacillus subtilis at 1.
Postgraduate Institute of Agricultur
De Vos, and O. Conformational changes in phosphoglucose isomerase induced by ligand binding.
Novel type of glucosephosphate isomerase in the hyperthermophilic archaeon Pyrococcus furiosus. Phylogenetic analyses and evolution of cPGIs. 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 Lateral gene transfers have been suggested for several conventional pgi genes 263144e.
Glucosephosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima: Generally, divalent metals were inhibitory at higher millimolar concentrations. The vials were then cooled on ice for 5 min, and the residual enzyme activity was tested and compared to the controls unheated samples.
The atmosphere is friendly and conducive to research. Pyrococcus furiosusSalmonella enterica serovar Typhimurium, and Ensifer meliloti.
Molecular, kinetic, thermophilic, and inhibitory properties were analyzed and compared. Journal List J Bacteriol v. In summary, we give an elaborate pbis of a novel protein family, the cPGI family, which represents a convergent line of PGIs.
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Support Center Support Center. Under these conditions activity remained about constant. Urbanke Hanover, Germany fprmat analytical ultracentrifugation and R. The region between the two cupin motifs, which varies from a minimum 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.
In order to obtain metal-free enzyme, 2 mg of enzyme was incubated in the presence of 0. Temperature optimum and thermostability.
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Thermococcales and Methanosarcinales Salmonella. Evolution of functional diversity in the cupin superfamily. Acknowledgments We thank C. Crystal structure of rabbit phosphoglucose isomerase, a glycolytic enzyme that moonlights as neuroleukin, autocrine motility factor, and differentiation mediator.
Indeed, in a phylogram including both cPGIs and several putative members of the cupin superfamily from eukaryal, pyis, and archaeal cPGIs form a separate cluster within the cupin superfamily not shown and thus represents its own cPGI family. Thus, under physiological conditions cPGIs presumably represent a heterogeneous mixture with respect to their metal content depending on both affinity and availability of the respective divalent metals. Comparison of molecular and kinetic properties of cPGIs from P.
The assumption of lateral gene transfers from a euryarchaeal source to some proteobacteria defines the cPGI family as an euryarchaeal family that originated during early euryarchaeal evolution. 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.
Additional putative homologs are currently not present in protein databases. Germin is a manganese containing homohexamer with oxalate oxidase and superoxide dismutase activities. Thus, the data presented here, and in particular the reactivation experiments, indicate an involvement of metals in catalysis.
Phosphoglucose isomerase from Escherichia coli K Identification, cloning and functional overexpression of cPGIs. Moreover, the presence thhesis 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