Novel type of glucosephosphate isomerase in the hyperthermophilic archaeon Pyrococcus furiosus. Thermostability of cPGIs from A. The latter assay was routinely used for most PGI activity determinations. Thus far this family comprises nine euryarchaeal and two proteobacterial sequences. The deduced activation energy of the latter was significant lower than the corresponding values of the other cPGIs in the present study. The lecture panel at PGIS well experienced and very helpful and they encourage the students to reach greater heights. The inserted gene sequences were confirmed on each strand by the method of Sanger et al.
Brandenburger is also gratefully acknowledged. PGI plays a central role in sugar metabolism of eukarya, bacteria, and archaea, both in glycolysis via the Embden-Meyerhof pathway in eukarya and bacteria and in its modified versions found in archaea. Erythrosephosphate and 6-phosphogluconate, classical inhibitors of the conventional PGIs with K i values in the micromolar range e. Low buffer concentrations 10 mM and Ultrapure Milli-Q water had to be used for all substances in the assays. Evolution of functional diversity in the cupin superfamily.
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Pgis thesis format outline – College Students Essay
Since the enzyme activities were not sensitive to oxygen, all assays were performed in the presence of oxygen. Koch for determination of the metal content.
Phylogenetic analyses and evolution of cPGIs. The measured K Me,0. The given pH values correspond to the respective temperature used.
Author information Article notes Copyright and License information Disclaimer. The deduced activation energy of the latter was significant lower than the corresponding values of the other cPGIs in the present study.
Phylogenetic analysis suggests lateral gene transfer for the bacterial cPGIs from euryarchaeota.
In summary, we give an elaborate characterization of a novel protein family, the cPGI family, which represents a convergent line of PGIs. Microbial relatives of fofmat seed storage proteins of higher plants: For the stcpgi gene, a lateral gene transfer has to be assumed after the lines of Formwt. Glucosephosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima: De Vos, and J.
Parameter Property a of: AKKUPROG was used to calculate the apparent molecular masses by fitting the ideal distribution for a single species to the measured concentration profiles. Crystal structure of rabbit phosphoglucose isomerase, a glycolytic enzyme that moonlights as neuroleukin, autocrine motility factor, and differentiation mediator. Tyr, Ala69, and Ala, which foemat been suggested to contribute to the hydrophobicity of the active site of PfcPGI, are not conserved.
The proposed origin of the cPGI family in the euryarchaeal evolution is indicated by an open circle, and the postulated lateral gene transfers are indicated by arrows.
Two distinct heterodisulfide reductase-like enzymes in the sulfate-reducing archaeon Archaeoglobus profundus. To prove their coding function as cpgithese ORFs were cloned and functionally overexpressed in Escherichia coli as follows. Sc in Computer Science.
In order to analyze the phylogenetic relation of cPGIs within the cupin superfamily, phylogenetic analyses formmat performed by two tree construction methods: This includes all proteobacteria, especially Agrobacterium tumefaciens and S. Erythrosephosphate and 6-phosphogluconate, classical inhibitors of the conventional PGIs with K i values in the micromolar range e.
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De Vos, and O. A total of 17 residues are completely conserved among these 11 sequences, including the metal-binding site His88, His90, His, and Glu97; PfcPGI numbering 430 All metals were used in the form of chlorides. The monomeric structure of the AfcPGI was verified by analytical ultracentrifugation. The partial specific volume of the protein was calculated from the amino acid sequence. Expression of the pfcpgi from the T7 promoter was more than five times higher than from the araC promoter see reference Thus far this family comprises nine euryarchaeal and two proteobacterial sequences.
Not surprisingly, cPGI family sequences cannot be divided into a bacterial and an euryarchaeal cluster but rather into two or three subfamilies: Structure of isopenicillin N synthase complexed with substrate and the mechanism of penicillin formation.
F6P, fructosephosphate; G6P, glucosephosphate. Temperature optimum and thermostability.