The first global screening of protein substrates bearing protein-bound 3,4-Dihydroxyphenylalanine in Escherichia coli and human mitochondria.
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.
Lee
Sangkyu
S
Ben May Department for Cancer Research, University of Chicago, Chicago, IL 60637, USA.
Chen
Yue
Y
Luo
Hao
H
Wu
Andrew A
AA
Wilde
Michael
M
Schumacker
Paul T
PT
Zhao
Yingming
Y
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
D015259:Dopamine Agents; D004295:Dihydroxyphenylalanine
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