Isolation and characterization of the 10-kDa and 22-kDa polypeptides of higher plant photosystem 2.
Two polypeptides of 10 kDa and 22 kDa, shown to be components of the higher plant photosystem 2, were purified and examined. A NaCl/Triton X-100 treatment was designed, which released these two polypeptides from the thylakoid membrane, in concert with the extrinsic 16-kDa and 23-kDa proteins, concomitant with a loss in oxygen-evolution activity. After this treatment the oxygen-evolving activity of the photosystem 2 membranes devoid of the 10-kDa and the 22-kDa polypeptides could be restored with CaCl2, but not by readdition of the purified 23-kDa protein. This deficiency was caused by an inability of the 23-kDa protein to rebind to the photosystem 2 membranes. In analogy, the oxygen-evolution activity of a highly purified photosystem 2 core preparation, devoid of the 10-kDa and 22-kDa polypeptides, was stimulated by CaCl2, but not by the 23-kDa protein. We, therefore, suggest that the 10-kDa or the 22-kDa polypeptides provide a binding-site for the extrinsic 23-kDa protein to the thylakoid membrane. The 10-kDa and 22-kDa polypeptides were isolated through ion-exchange chromatography in the presence of detergents. They both displayed hydrophobic properties, verified by their low proportion of polar amino acid residues and their partition to the hydrophobic phase during Triton X-114 fractionation. The purified polypeptides did not contain metallic cofactors or substances with absorption in the visible region of the spectrum.
D000596:Amino Acids / Q000032:analysis; D002122:Calcium Chloride / Q000494:pharmacology; D002734:Chlorophyll / Q000032:analysis*; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D003429:Cross Reactions; D045342:Light-Harvesting Protein Complexes; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins / Q000032:analysis*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology
D000596:Amino Acids; D002793:Cholic Acids; D045342:Light-Harvesting Protein Complexes; D010455:Peptides; D045322:Photosynthetic Reaction Center Complex Proteins; D010940:Plant Proteins; D002734:Chlorophyll; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D019826:Cholic Acid; D002122:Calcium Chloride