Contribution to the structural characterization of eucaryotic PSI reaction centre-I. Critical analysis of the polypeptidic composition of different P700 enriched fractions.
In thylakoid membranes, several peptides of high MW† are present which may interfere with the study of CP1's components. Modifying Cleveland's technique [7] for limited proteolysis, we have characterized the polypeptides found in the 60 kD region. Some may result from incomplete washing of the CF1 while others come from the CP1; indeed, this chlorophyll protein complex, which has a higher MW (above 100 kD), very often undergoes a dissociation into smaller components of about 60 KD MW.Analysis of the protein content of different preparations commonly used to obtain PSI reaction centre enriched fractions has been performed. The α and β subunits of CF1 are among the main contaminants of most of these preparations. A further purification step is described which can be applied to all these preparations, but numerous peptides are still present in the active fractions. It is most unlikely that all these polypeptides are required for the primary photochemical event, and this emphasizes the necessity to find a new simple method to purify PSI reaction centres.
Service de Biochimie, Departement de Biologie, C.E.N. de SACLAY, BP no2, Gif-sur-Yvette, 91 190, France.
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