When higher buffer concentrations or deionized H 2 O was used, the residual catalytic activity was still measured data not shown after EDTA treatment, which was presumably due to soaking of metal impurities into the metal-binding site of the enzymes. Additional putative homologs are currently not present in protein databases. New Year Celebrations – Author information Article notes Copyright and License information Disclaimer. Journal List J Bacteriol v.

Sequence comparison and phylogenetic affiliation. I really valued the support and encouragement received from lecturers and my colleagues. Spectra were recorded before and after each temperature gradient. The following controls were used. EDTA, erythrosephosphate, 6-phosphogluconate, sorbitolphosphate, phosphoenolpyruvate, ADP, fructosephosphate, and fructose-1,6-bisphosphate. Structure of native phosphoglucose isomerase from rabbit: Transkingdom transfer of the phosphoglucose isomerase gene.

Kinetic data were fit thess the Michaelis-Menten equation with the MicroCal software by using the Levenberg-Marquard algorithm. 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 topology was achieved by both methods NJ and ML and is supported by very good bootstrapping values.

pgis thesis format

Support Center Support Center. A scenario would be plausible in which these novel cPGIs have replaced an ancestral conventional euryarchaeal PGI before the Thermococcales evolved.

Sc in Applied Statistics. Loss of metal-binding ability in some TlcPGI mutants goes along with a loss of catalytic function, suggesting a role in catalysis PGI activity containing fractions were recovered from the last three elution fractions, the one obtained from the last two were essentially pure.


Postgraduate Institute of Agricultur

Accession numbers were as follows: Janani de Silva M. The deduced activation energy of the latter was significant lower than the corresponding values of the other cPGIs in the present study.

Thus far, conclusive biochemical studies analyzing the role of metals have not been performed. Structure of isopenicillin N synthase complexed with substrate and the mechanism of penicillin formation.

Koch for determination of the metal content.

Postgraduate Institute of Science

Sc in Computer Science at PGIS I have enhanced my career prospects and gained additional knowledge which is very helpful in my career as a lecturer. Protein concentrations were determined by the method of Bradford 5 with bovine serum formzt as a standard.

A rapid and sensitive method for the quantitation of microgram quantities of protein pvis the principle of protein-dye binding. Their presence is presumably the result of a lateral gene transfer from a Thermococcus -like species after the lines of E. Phylogenetic analyses suggest that bacterial cPGIs originated from euryarchaeota by lateral gene transfer. University of Hanover, Hanover, Germany.

AKKUPROG was used to calculate the apparent molecular masses by fitting the ideal distribution for a single species to the measured concentration profiles.

Pgis thesis format outline

Thermal unfolding of cPGIs from P. Novel type of glucosephosphate isomerase in the hyperthermophilic archaeon Pyrococcus furiosus.


pgis thesis format

Programmes – January. Secondary structure analysis and assignment to different secondary structure types were performed by the experimentally established spectra-structure correlation by using the Varselec option of Dicroprot 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 The given pH values correspond to the respective temperature used. In most cases the corresponding metal-binding site of forrmat cupin superfamily proteins is coordinated by three histidines and a glutamate as well F6P, fructosephosphate; G6P, glucosephosphate.

All metals were used in thessi form of chlorides.

Thus, the evolution and distribution of PGIs in the phylogenetic tree is complex and rather puzzling Fig. DNA sequencing with chain-terminating rormat. This includes all proteobacteria, especially Agrobacterium tumefaciens and S.

Confidence limits were estimated by bootstrapping replicates. In this report we give a comprehensive characterization of cPGIs from archaea and bacteria which represent a novel PGI family.