The carboxyl-terminal tripeptide of the manganese-stabilizing protein is required for quantitative assembly into photosystem II and for high rates of oxygen evolution activity.
The extrinsic manganese stabilizing protein of photosystem II is required for Mn retention by the O2-evolving complex, accelerates the rate of O2 evolution, and protects photosytem II against photoinhibition. We report results from studies of the in vitro reconstitution of spinach photosytem II with recombinant manganese stabilizing protein with C-terminal deletions of two, three, and four amino acids. The deletions were the result of amber mutations introduced by site-directed mutagenesis. Removal of the C-terminal dipeptide (Glu-Gln) did not diminish the ability of the manganese stabilizing protein either to rebind to or to restore high rates of O2 evolution to photosystem II preparations depleted of the native protein. Deletion of the C-terminal tripeptide (Leu-Glu-Gln) resulted in weakened but specific binding of manganese stabilizing protein to photosystem II and minimal recovery of O2 evolution activity. Removal of the C-terminal tetrapeptide (Gln-Leu-Glu-Gln) eliminated the ability of the subunit to interact stably with all of its available binding sites on photosystem II, as evidenced by the fact that this mutant was totally inactive in restoring O2 evolution activity. Evidence is presented to indicate that these mutational effects on the binding and function of the manganese stabilizing protein may be due to major changes in tertiary structure. The truncation mutations lacking either the C-terminal tri- or tetrapeptide exhibit apparent size increases of 25 and 40%, respectively, when compared either to a mutant lacking the C-terminal dipeptide or to the wild-type protein.
Department of Biology, University of Michigan, Ann Arbor 48109-1048, USA.
S D
SD
Betts
N
N
Lydakis-Simantiris
J R
JR
Ross
C F
CF
Yocum
D017360:Arabidopsis; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics; D005973:Glutamine / Q000235:genetics; D007930:Leucine / Q000235:genetics; D008345:Manganese / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D018724:Spinacia oleracea
D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D011506:Proteins; D011994:Recombinant Proteins; C064388:oxygen-evolving complex, 33 kDa protein; C054266:photosystem II manganese-stabilizing protein; D005973:Glutamine; D008345:Manganese; D007930:Leucine; D010100:Oxygen