The main aim of the project is the further development of the applicant's skills and expertise in the application of the surface plasmon resonance imaging technique (SPR) for the investigation of interactions of proteins and other bioactive compounds.
The project consists of two particular targets:
i) the introduction of the SPR imaging technique to the host laboratory (University in Bialystok, Poland) in order to enhance host research potential in the area of interactions between cholesterol, phospholipids and cellular membrane proteins,
ii) development of a sensor for different cathepsins. Cholesterol is the major component of cell membranes. It also plays a crucial role in arteriosclerosis development.
The construction of a sensor with cholesterol immobilized on the sensor surface has potential for the investigation of cholesterol- protein interactions. Cholesterol mono-succilic acid will be immobilized to the thiol first layer on the gold chip. The immobilization will be performed with EDC and NHS application. Cathepsins are enzymes - lysosomal cysteine proteases, which play an important role in protein degradation, antigen presentation, bone resorption and hormone processing. These enzymes have also been implicated in tumour invasion and methastasis. Developing a sensor for cathepsins is considered ideal for the application of the SPR imaging system.
In order to develop a sensor for different cathepsins, the strong interaction of cathepsins with cystatin will be used. Cystatin C seems to be the most suitable for this purpose. In order to immobilize cystaine C, different thiols will be investigated as the first layer on gold. The active group of immobilized thiol is the carboxy group or the amine group. These groups react with protein and create covalent bonds. The linear Surface Plasmon Resonance system will be used for measurements consisting of a He-Ne laser, two polarizators, and two lenses for obtaining polarization light.
Fields of science
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics
- engineering and technologymaterials engineering
- natural scienceschemical sciencesinorganic chemistrytransition metals
- natural sciencesphysical sciencesopticslaser physics
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
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