Evidence for a post-translational modification, aspartyl aldehyde, in a photosynthetic membrane protein.
In oxygenic photosynthesis, photosystem II (PSII) carries out the oxidation of water and reduction of plastoquinone. Three PSII subunits contain reactive groups that covalently bind amines and phenylhydrazine. It has been proposed that these reactive groups are carbonyl-containing, co- or post-translationally modified amino acids. To identify modified amino acid residues in one of the PSII subunits (CP47), tandem mass spectrometry was performed. Modified residues were affinity-tagged with either biotin-LC-hydrazide or biocytin hydrazide, which are known to label carbonyl groups. The affinity-tagged subunit was isolated by denaturing gel electrophoresis, and tryptic peptides were then subjected to affinity purification and tandem mass spectrometry. This procedure identified a hydrazide-labeled peptide, which has the sequence XKEGR. This result is supported by quantitative results acquired from peptide mapping and methylamine labeling. The gene sequence and these tandem data predict that the first amino acid, X, which is labeled with the hydrazide reagent, is a modified form of aspartic acid. On the basis of these data, we propose that D348 of the CP47 subunit is post- or co-translationally modified to give a novel amino acid side chain, aspartyl aldehyde.
Anderson
Lorraine B
LB
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, St. Paul, Minnesota 55108, USA.
Ouellette
Anthony J A
AJ
Eaton-Rye
Julian
J
Maderia
Melissa
M
MacCoss
Michael J
MJ
Yates
John R
JR
Barry
Bridgette A
BA
D000596:Amino Acids / Q000032:analysis; D045342:Light-Harvesting Protein Complexes / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D045332:Photosystem II Protein Complex / Q000737:chemistry* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021122:Protein Subunits; D018724:Spinacia oleracea / Q000737:chemistry / Q000378:metabolism
D000596:Amino Acids; D045342:Light-Harvesting Protein Complexes; D045332:Photosystem II Protein Complex; D021122:Protein Subunits; C070970:photosystem II, chlorophyll-binding protein, CP-47