An activated glutamate residue identified in photosystem II at the interface between the manganese-stabilizing subunit and the D2 polypeptide.
Photosystem II (PSII) catalyzes the oxidation of water during oxygenic photosynthesis. PSII is composed both of intrinsic subunits, such as D1, D2, and CP47, and extrinsic subunits, such as the manganese-stabilizing subunit (MSP). Previous work has shown that amines covalently bind to amino acid residues in the CP47, D1, and D2 subunits of plant and cyanobacterial PSII, and that these covalent reactions are prevented by the addition of chloride in plant preparations depleted of the 18- and 24-kDa extrinsic subunits. It has been proposed that these reactive groups are carbonyl-containing, post-translationally modified amino acid side chains (Ouellette, A. J. A., Anderson, L. B., and Barry, B. A. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 2204-2209 and Anderson, L. B., Ouellette, A. J. A., and Barry, B. A. (2000) J. Biol. Chem. 275, 4920-4927). To identify the amino acid binding site in the spinach D2 subunit, we have employed a biotin-amine labeling reagent, which can be used in conjunction with avidin affinity chromatography to purify biotinylated peptides from the PSII complex. Multidimensional chromato-graphic separation and multistage mass spectrometry localizes a novel post-translational modification in the D2 subunit to glutamate 303. We propose that this glutamate is activated for amine reaction by post-translational modification. Because the modified glutamate is located at a contact site between the D2 and manganese-stabilizing subunits, we suggest that the modification is important in vivo in stabilizing the interaction between these two PSII subunits. Consistent with this conclusion, mutations at the modified glutamate alter the steady-state rate of photosynthetic oxygen evolution.
School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
Sascha
S
Rexroth
Catherine C L
CC
Wong
Jessica H
JH
Park
John R
JR
Yates
Bridgette A
BA
Barry
D001665:Binding Sites; D005075:Biological Evolution; D002264:Carboxylic Acids / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D018698:Glutamic Acid / Q000737:chemistry*; D008345:Manganese / Q000737:chemistry*; D013058:Mass Spectrometry; D008968:Molecular Conformation; D009154:Mutation; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D010788:Photosynthesis; D045332:Photosystem II Protein Complex / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D014443:Tyrosine / Q000737:chemistry
D002264:Carboxylic Acids; D045332:Photosystem II Protein Complex; D018698:Glutamic Acid; D014443:Tyrosine; D008345:Manganese; D010100:Oxygen