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Content archived on 2024-05-29

NMR studies of protein structures

Objective

The outgoing phase: Structural proteomics is a large-scale project to determine a sufficiently large number of structures to be representative of all protein folds. This objective is approached using high-throughput protein preparation and structure determination with X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy. My particular interest involves the use of NMR spectroscopy in structural proteomics.

The development of NMR spectroscopy for structural proteomics of Thermotoga maritima is a major focus of the research group of Professor Kurt Wüthrich at The Scripps Research Institute, which forms a part of the Joint Centre for Structural Genomics.

About 60% of T. maritima proteins do not have predicted structural annotations, and their levels of homology to proteins with known structures are lower than 30%. The structurally unpredictable proteins with fewer than 150 amino acid residues will be targets for my structural studies. The structures of proteins exhibiting promising NMR spectra will be determined using NMR spectroscopy. The determined structures will provide a basis for further functional studies. The return phase: Enzymes are of great interest to chemical and pharmaceutical industry as asymmetric catalysts. Humicola lanuginosa lipase (HLL) hydrolyses water-insoluble esters, and is applied as a detergent constituent. The proposed research deals with the studies on structural basis of HLL enantioselectivity.

The enzyme will be in vitro evolved towards improved enantioselectivity by a number of cycles involving:
i) introduction of mutations, and
ii) selection of improved HLL using high-throughput screening method.

The structure of the evolved protein will be studied with NMR spectroscopy. Comparison of the obtained structure with the wild type will reveal structural basis of HLL enantioselectivity.

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Keywords

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Topic(s)

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Call for proposal

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FP6-2004-MOBILITY-6
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Funding Scheme

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OIF - Marie Curie actions-Outgoing International Fellowships

Coordinator

MAX-PLANCK-INSTITUT FUR KOHLENFORSCHUNG
EU contribution
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Total cost

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Participants (1)

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