Quantitative Analysis of Genetically Altered Reaction Centers Using an In Vitro Cytochrome Oxidation Assay
In the analysis of complex phenomena such as the biogenesis or photochemical function of reaction center proteins, the approach of a geneticist is to generate and characterize a large number of mutants affecting specific steps in the process. In some analyses, spontaneous mutants which possess selectable characteristics may be informative. For example, the role of specific amino acid residues in herbicide and quinone binding can be deduced from the analysis of spontaneous herbicide resistant mutants. However, the analysis of spontaneous mutations as a general approach to study structure-function relationships has serious limitations. Many mutations have phenotypes which possess unselectable characteristics. With the determination of the structure of the reaction center1, this class of mutations can be generated by replacing important amino acid residues adjacent to chromophores throughout the protein by site-directed mutagenesis.
KeywordsReaction Center Reaction Center Protein Flash Intensity Relative Light Intensity Multiple Photon
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