Research objectives and content
The aim of the present proposal is to determine the role played by a number of specific amino acids of the cyanobacterial cytochrome c6, which are presumably important for its fuction as electron donor of photosystem I. Special attention should be payed to those residues which participate in either molecular recognition (by formation of electrostatic interaction between the two redox partners) or electron transfer.
A structural model of Anabaena cyt c6 should be made by energy minimization and molecular dynamics calculations, after aligning the residue sequence to that of cyt c6 from the eukaryotic alga M. braunii, for which the structure has recently been solved. The model should be used both for designing site directed mutants and for checking possible structural changes due to modifications. In order to test the functionality of modified proteins, a kinetic analysis of electron transfer from wild-type and mutant cytochrome c6 to PSI should be carried out by laser flash spectroscopy.
Training content (objective, benefit and expected impact)
The applicant should profite from the expertise of the host group on fast kinetic spectroscopy and molecular biology techniques while developing his expertise on theoretical calculations. His training on structural techniques acquired during his postdoctoral stays will be highly useful for the host group. As a result of the proposed research, our understanding of the structure-function relationships and molecular recognition processes in redox proteins will be significantly increased