Objective
Characterization of a range of triosephophate isomerases (TIMs) which differ widely in stability.
Creation of monomeric mutants of TIM by either site directed mutagenesis or complexation with tight binding cyclopeptides, or via both approaches.
Eventually, with the help of model building techniques, the implementation of loop transplantations will be attempted in such a way that single monomers are formed with an altered specificity and/or new catalytic activity.
The current objectives involve research on: trypanosomal TIM; human TIM; Escherichia coli TIM; octarellin design; psychrophilic TIM and thermophilic TIM; cyclopeptides; and loop transplantation modelling studies.
Major results to date include: TIM variants of trypanosomal TIM and E coli TIM have been made and partly characterized;
the sequences of TIM from thermophilic Bacillus Stearothermophilus and psychrotrophic Moraxella spp TA137 have been determined;
the structural change from an oxopeptide to the thiopeptide cyclo(Pheomega [CSNH]-Phe-Gly-Pro-Phe-Val-) results in a drastic increase in biological activity;
the crystal structure of wild type E coli TIM has been determined at 2.6 angstroms resolution.
A network of European research groups has been formed to study and engineer the stability and activity of the dimeric enzyme triosephosphate isomerase (TIM) with the ultimate goal of being able to create new classes of enzymes based on the (beta alpha)8-framework.
Within this collaboration psychrotrophic and thermophylic TIMs will be studied, including the crystal structure determinations. This data will help to evaluate important characteristics related to the stability of this enzyme. On the basis of the refined crystal structure of trypanosomal TIM, mutants will be made in which the dimer interface interactions are weakened. The eventual goal is to obtain monomeric TIM (complexed with or without cyclopeptides which mimic the dimer interface loops) with an altered specificity and/or new catalytic activity.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- engineering and technology materials engineering crystals
- medical and health sciences clinical medicine transplantation
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Coordinator
HEIDELBERG
Germany
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