Structural biology: Structural and functional characterisation of two myoglobins with different functions.
Myoglobin is the protein responsible for oxygen storage and transport in heart and skeletal muscle, however, some organisms have two similar myoglobins that functions differently in different tissues, one with main function to store oxygen, while the other to detoxify peroxides and other reactive species. The focus will be to elucidate the structural basis governing this functional difference. This is a master project in biochemistry and structural biology.
Myoglobin (Mb) has been extensively studied for more than a hundred years, and it was the first protein of which the structure was solved back in the 1950ties. Recently, it has been shown that Mb can also have other functions than just oxygen storage and transport. It can scavenge reactive oxygen species like peroxides and thereby protecting the heart, or remove or produce the signaling molecule nitric oxide that can inhibit the mitochondrial respiration. We have previously studied the reaction with peroxides in horse heart Mb. Now it has been discovered that common carp contains two Mbs, one that is expressed in several tissues Mb1 and one exclusively in brain tissue Mb2. The Mb1 has been shown to better bind oxygen than Mb2, while Mb2 reacts more efficiently with peroxides. This master project will focus on trying to understand the structural basis that governs this functional difference.
The project will involve establishing a purification procedure for these two proteins, crystallise them and solve the structures with X-Ray diffraction / protein crystallography. A further aspect will be to optimise crystallisation for growing large crystals, that can be used for neutron diffraction. Different structures will be solved with bound oxygen and reacted with peroxide to be able to understand their differences. Data collect for solving the structures will involve travelling abroad to synchrotrons. Additionally, the structural studies will be supplemented by some spectroscopic characterisation. The biophysical and biochemical methods used in this master project will to a large extend be covered in BIOS4020.
Methods you will use and learn:
- Protein expression
- Protein purification
- Protein crystallisation
- Solving the crystal structure of proteins
- Enzyme kinetics
- Redox potential measurements
- Anaerobic work
- Spectroscopic charaterisation.
Data collection for solving the structures will involve travelling abroad to synchrotrons. The biophysical and biochemical methods used in this master project will to a large extend be covered in BIOS4020, taught by the supervisors.
Through the master project, you will also learn to
- Present your work orally through group meetings
- Present your work through posters presentations at scientific conferences.
- Learn to plan and perform scientific work
- Learn to write up your work as a thesis
Supervision: The master project will be performed in the Structural Redox Biochemistry - Hersleth Group (Section for Biochemistry and Molecular Biology) and supervised by Hans-Petter Hersleth and Marta Hammerstad.
Contact: Hans-Petter Hersleth room 2313, e-mail: firstname.lastname@example.org
To read more see our group homepage: http://hersleth.org/