Biophysical aspects of de novo designed metalloproteins

– Course content:  This course will combine theoretical and practical instruction to explore the field of de novo designed proteins tailored to contain metal redox cofactors. There will be theoretical lectures and demonstrations/hands-on laboratory experiences. The lectures will describe the factors that must be considered in order to generate stable assemblies of small designed peptides; how to synthesize, purify and characterize synthetic peptides; and methods to engineer metal binding sites within these constructs. The practical lab work will focus on samples preparation and characterization using UV-Vis electronic absorption and 9-GHz Electron Paramagnetic Resonance (EPR) spectroscopies of synthetic mini-proteins that bind hemin within the coiled-coil scaffold.

Course Outcomes: The student that completes this course will gain a fundamental understanding of:

1) the factors that lead to proper protein folding, especially for a-helical proteins.

2) strategies to design a-helical self-assembling coiled coils.

3) approaches to engineer metal binding sites into designed scaffolds.

4) methods to characterize designed proteins using spectroscopic techniques such as UV-Vis electronic absorption and EPR spectroscopies.

Timeline

– Timeline: 24, 25 and 26 March 2025

Students prerequisites

• Basic knowledge of Chemistry/Bioinorganic Chemistry, Biochemistry.
• Agreement of the thesis director
• The prior reading of references (ref 3 below is mandatoryfor a good ouput from the lab sessions given the relativey short time. Similarly, reading of references (1) and (2) will be beneficial to get a better understanding of the theoretical lectures.