The first global screening of protein substrates bearing protein-bound 3,4-dihydroxyphenylalanine in escherichia coli and human mitochondria

Sangkyu Lee, Yue Chen, Hao Luo, Andrew A. Wu, Michael Wilde, Paul T. Schumacker, Yingming Zhao

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)5705-5714
Number of pages10
JournalJournal of Proteome Research
Volume9
Issue number11
DOIs
StatePublished - Nov 5 2010

Fingerprint

Bearings (structural)
Dihydroxyphenylalanine
Mitochondria
Escherichia coli
Screening
Substrates
Phenylalanine
Proteins
Hydroxylation
Intrinsically Disordered Proteins
Glyceraldehyde-3-Phosphate Dehydrogenases

Keywords

  • Escherichia coli
  • PB-DOPA
  • Protein hydroxylation
  • human mitochondria
  • protein-bound 3,4-dihydroxy-phenylalanine
  • tyrosine oxidation

Cite this

The first global screening of protein substrates bearing protein-bound 3,4-dihydroxyphenylalanine in escherichia coli and human mitochondria. / Lee, Sangkyu; Chen, Yue; Luo, Hao; Wu, Andrew A.; Wilde, Michael; Schumacker, Paul T.; Zhao, Yingming.

In: Journal of Proteome Research, Vol. 9, No. 11, 05.11.2010, p. 5705-5714.

Research output: Contribution to journalArticle

Lee, Sangkyu ; Chen, Yue ; Luo, Hao ; Wu, Andrew A. ; Wilde, Michael ; Schumacker, Paul T. ; Zhao, Yingming. / The first global screening of protein substrates bearing protein-bound 3,4-dihydroxyphenylalanine in escherichia coli and human mitochondria. In: Journal of Proteome Research. 2010 ; Vol. 9, No. 11. pp. 5705-5714.
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