Abstract
This chapter describes biochemical and molecular genetic methods and resources to study photosystem I (PSI) of Synechocystis sp. PCC 6803. In recent years, cyanobacteria have been used increasingly to study structure-function relations in photosynthetic proteins, including PSI. The mesophilic cyanobacterium, Synechocystis sp. PCC 6803 is a model system for using molecular genetic approaches to study functions of PSI proteins. Its genome is completely sequenced and the protein components of its photosystem are identified. Targeted mutations in all genes for PSI proteins are available. To study the topography of PSI, it is important to probe the exposed protein surface by modification of specific residues. Two biotin derivatives can be used to modify surface-exposed residue. N-hydroxysuccinimidobiotin (NHS-biotin) specifically reacts with the N terminus and the e-amino group of lysyl residues, while biotin-maleimide (M-biotin) modifies the sulfhydryl groups of cysteinyl residues. Both reagents are used to study surface-exposed residues in the PSI complex.
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