Carboxylate groups on the manganese-stabilizing protein are required for its efficient binding to photosystem II.
The effects of the modification of carboxylate groups on the manganese-stabilizing protein of photosystem II were investigated. Carboxylate groups (including possibly the C-terminus) on the manganese-stabilizing protein were modified with glycine methyl ester in a reaction facilitated by 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide. The manganese-stabilizing protein that was modified while associated with NaCl-washed photosystem II membranes contained 1-2 modified carboxylates, whereas the protein that was modified while free in solution contained 4 modified carboxylates. Both types of modified protein could reconstitute oxygen evolution at high manganese-stabilizing protein to photosystem II reaction center ratios. However, the protein that had been modified in solution exhibited a dramatically altered binding affinity for photosystem II. No such alteration in binding affinity was observed for the protein that had been modified while associated with the photosystem. Mapping of the sites of modification was carried out by trypsin and Staphylococcus V8 protease digestion of the modified proteins and analysis by matrix-assisted laser desorption/ionization mass spectrometry. These studies indicated that the domains (157)D-(168)D and (212)E-(247)Q (C-terminus) are labeled only when the manganese-stabilizing protein is modified in solution. Modified carboxylates in these domains are responsible for the altered binding affinity of this protein for the photosystem.
Frankel
L K
LK
Department of Biological Sciences, Biochemistry and Molecular Biology Section, Louisiana State University, Baton Rouge 70803, USA.
Cruz
J A
JA
Bricker
T M
TM
D000595:Amino Acid Sequence; D001665:Binding Sites; D002264:Carboxylic Acids / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D007700:Kinetics; D008345:Manganese / Q000378:metabolism*; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism; D010449:Peptide Mapping; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018724:Spinacia oleracea
D002264:Carboxylic Acids; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D011506:Proteins; C054266:photosystem II manganese-stabilizing protein; D008345:Manganese; C035591:glycine methyl ester; D005022:Ethyldimethylaminopropyl Carbodiimide; D010100:Oxygen; D005998:Glycine