title: The first global screening of protein substrates bearing protein-bound 3,4-Dihydroxyphenylalanine in Escherichia coli and human mitochondria.
abstract: Protein hydroxylation at proline and lysine residues is known to have important effects on cellular functions, such as the response to hypoxia. However, protein hydroxylation at tyrosine residues (called protein-bound 3,4-dihydroxy-phenylalanine (PB-DOPA)) has not been carefully examined. Here we report the first proteomics screening of the PB-DOPA protein substrates and their sites in Escherichia coli and human mitochondria by nano-liquid chromatography/tandem mass spectrometry (nano-LC/MS/MS) and protein sequence alignment using the PTMap algorithm. Our study identified 67 novel PB-DOPA sites in 43 E. coli proteins and 9 novel PB-DOPA sites in 7 proteins from HeLa mitochondria. Bioinformatics analysis indicates that the structured region is more favored than the unstructured regions of proteins for the PB-DOPA modification. The PB-DOPA substrates in E. coli were dominantly enriched in proteins associated with carbohydrate metabolism. Our study showed that PB-DOPA may be involved in regulation of the specific activity of certain evolutionarily conserved proteins such as superoxide dismutase and glyceraldehyde 3-phosphate dehydrogenase, suggesting the conserved nature of the modification among distant biological species. The substrate proteins identified in this study offer a rich source for determining their regulatory enzymes and for further characterization of the possible contributions of this modification to cellular physiology and human diseases.
authors.affiliation: Ben May Department for Cancer Research, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Sangkyu
authors.initials: S
authors.lastname: Lee
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Hao
authors.initials: H
authors.lastname: Luo
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Wu
authors.forename: Michael
authors.initials: M
authors.lastname: Wilde
authors.forename: Paul T
authors.initials: PT
authors.lastname: Schumacker
authors.forename: Yingming
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D001665:Binding Sites; D050260:Carbohydrate Metabolism; D004295:Dihydroxyphenylalanine / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D015259:Dopamine Agents; D004926:Escherichia coli / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity
chemical_list: D015259:Dopamine Agents; D004295:Dihydroxyphenylalanine
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