EVIDENCE FOR A POST-TRANSLATIONAL MODIFICATION, ASPARTYL ALDEHYDE

天冬酰醛翻译后修饰的证据

• 批准号: 7723633
• 负责人: BRIDGETTE ANNE BARRY
• 金额: $ 0.08万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723633
• 关键词: Affinity; Affinity Chromatography; Aldehydes; Amines; Amino Acids; Aspartic Acid; Binding; Biotin; Computer Retrieval of Information on Scientific Projects Database; Data; Databases; Funding; Genes; Grant; Institution; Label; Methylamines; Peptide Mapping; Peptides; Phenylhydrazines; Photosynthesis; Plastoquinone; Post-Translational Protein Processing; Procedures; Reagent; Research; Research Personnel; Resources; Side; Source; United States National Institutes of Health; Water; biocytin hydrazide; carbonyl group; gel electrophoresis; methylamine; novel; oxidation; phenylhydrazine; photosystem II; tandem mass spectrometry

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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 (Ouellette et al. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 2204 and Anderson et al. J. Biol. Chem. 2000, 275, 4920). 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.

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