The oxidation state of the photosystem II manganese cluster influences the structure of manganese stabilizing protein.
Exposure of photosystem II membranes to trypsin that has been treated to inhibit chymotrypsin activity produces limited hydrolysis of manganese stabilizing protein. Exposure to chymotrypsin under the same conditions yields substantial digestion of the protein. Further probing of the unusual insensitivity of manganese stabilizing protein to trypsin hydrolysis reveals that increasing the temperature from 4 to 25 degrees C will cause some acceleration in the rate of proteolysis. However, addition of low (100 microM) concentrations of NH2OH, that are sufficient to reduce, but not destroy, the photosystem II Mn cluster, causes a change in PS II-bound manganese stabilizing protein that causes it to be rapidly digested by trypsin. Immunoblot analyses with polyclonal antibodies directed against the N-terminus of the protein, or against the entire sequence show that trypsin cleavage produces two distinct peptide fragments estimated to be in the 17-20 kDa range, consistent with proposals that there are 2 mol of the protein/mol photosystem II. The correlation of trypsin sensitivity with Mn redox state(s) in photosystem II suggest that manganese stabilizing protein may interact either directly with Mn, or alternatively, that the polypeptide is bound to another protein of the photosystem II reaction center that is intimately involved in binding and redox activity of Mn.
Hong
S K
SK
Department of Biology and Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1048, USA.
Pawlikowski
S A
SA
Vander Meulen
K A
KA
Yocum
C F
CF
D064753:Ammonium Hydroxide; D002918:Chymotrypsin; D004591:Electrophoresis, Polyacrylamide Gel; D006878:Hydroxides; D015151:Immunoblotting; D008345:Manganese / Q000737:chemistry*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D013696:Temperature; D014357:Trypsin
D006878:Hydroxides; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D011506:Proteins; C054266:photosystem II manganese-stabilizing protein; D008345:Manganese; D064753:Ammonium Hydroxide; D002918:Chymotrypsin; D014357:Trypsin; D010100:Oxygen