Final Activity Report Summary - NMRSTRUCTPROT (NMR studies of protein structures)
Nuclear magnetic resonance (NMR) is a tool used to determine the three-dimensional structure of a protein. NMR provides additional information about protein dynamics. This data can then be related to protein activity, such as interactions with drugs or catalytic properties.
The outgoing phase project was part of the structural proteomics initiative, which aimed to determine all unknown protein folds, thus providing a large testing set for structure prediction tool development. Structures of two proteomics targets were determined using nuclear magnetic resonance (NMR) and two other structure determinations were also attempted.
A protein selection procedure suitable for structure determination was developed. Modern equipment, namely a 1 mm diameter probe combined with robotics, allowed for high-throughput screening of proteins using low amounts of material. In addition, NMR screening was established as a tool to select folded proteins amenable for structure determination using X-ray crystallography.
The return phase project aimed to determine factors responsible for protein thermostability. The thermal stability of proteins is of great importance in biotechnology and industry, where it is desired to use stable and robust catalysts. Lipase A from bacillus subtilis was used as a model protein for this study. Comparison of the structure and properties between two variants of the protein was performed. The mutants differed at only one amino-acid position which induced change in thermostability of 30 degrees Celsius. The studies were performed using NMR spectroscopy and X-ray crystallography and were in progress at the reporting date.
The outgoing phase project was part of the structural proteomics initiative, which aimed to determine all unknown protein folds, thus providing a large testing set for structure prediction tool development. Structures of two proteomics targets were determined using nuclear magnetic resonance (NMR) and two other structure determinations were also attempted.
A protein selection procedure suitable for structure determination was developed. Modern equipment, namely a 1 mm diameter probe combined with robotics, allowed for high-throughput screening of proteins using low amounts of material. In addition, NMR screening was established as a tool to select folded proteins amenable for structure determination using X-ray crystallography.
The return phase project aimed to determine factors responsible for protein thermostability. The thermal stability of proteins is of great importance in biotechnology and industry, where it is desired to use stable and robust catalysts. Lipase A from bacillus subtilis was used as a model protein for this study. Comparison of the structure and properties between two variants of the protein was performed. The mutants differed at only one amino-acid position which induced change in thermostability of 30 degrees Celsius. The studies were performed using NMR spectroscopy and X-ray crystallography and were in progress at the reporting date.