Biosynthesis and turnover of DOPA-containing proteins by human cells

Kenneth Rodgers, Peter Hume, Rachael Dunlop, Roger Dean

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Protein-bound 3,4-dihydroxyphenylalanine (PB-DOPA) is a major product of hydroxyl radical attack on tyrosine residues of proteins. Levels of PB-DOPA in cells and tissues have been shown to be greatly elevated in age-related diseases. We demonstrate for the first time that l-DOPA (levodopa) can be biosynthetically incorporated into cell proteins by human cells (THP-1 monocytes and monocyte-derived macrophages). The DOPA-containing proteins generated were selectively visualized on PVDF membranes using a redox-cycling staining method. Many cell proteins contained DOPA and seemed to be synthesized as their full-length forms. The cellular removal of DOPA-containing proteins by THP-1 cells was by proteolysis involving both the proteasomal and the lysosomal systems. The rate of cellular proteolysis of DOPA-containing proteins increased at lower levels of DOPA incorporation but decreased at higher levels of DOPA incorporation. The decreased rate of degradation was accompanied by an increase in the activity of cathepsins B and L but the activity of cathepsin S increased only at lower levels of DOPA incorporation. These data raise the possibility that PB-DOPA could be generated in vivo from l-DOPA, which is the most widely used treatment for Parkinson disease.

Original languageEnglish
Pages (from-to)1756-1764
Number of pages9
JournalFree Radical Biology Medicine
Volume37
Issue number11
DOIs
Publication statusPublished - 2004

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Biosynthesis
Cells
Dihydroxyphenylalanine
Proteins
Proteolysis
cathepsin S
Cathepsin L
Cathepsin B
Macrophages
Levodopa
Hydroxyl Radical
Oxidation-Reduction
Parkinson Disease
Tyrosine
Monocytes
Tissue
Staining and Labeling
Membranes
Degradation

Cite this

Rodgers, Kenneth ; Hume, Peter ; Dunlop, Rachael ; Dean, Roger. / Biosynthesis and turnover of DOPA-containing proteins by human cells. In: Free Radical Biology Medicine. 2004 ; Vol. 37, No. 11. pp. 1756-1764.
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Biosynthesis and turnover of DOPA-containing proteins by human cells. / Rodgers, Kenneth; Hume, Peter; Dunlop, Rachael; Dean, Roger.

In: Free Radical Biology Medicine, Vol. 37, No. 11, 2004, p. 1756-1764.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Biosynthesis and turnover of DOPA-containing proteins by human cells

AU - Rodgers, Kenneth

AU - Hume, Peter

AU - Dunlop, Rachael

AU - Dean, Roger

PY - 2004

Y1 - 2004

N2 - Protein-bound 3,4-dihydroxyphenylalanine (PB-DOPA) is a major product of hydroxyl radical attack on tyrosine residues of proteins. Levels of PB-DOPA in cells and tissues have been shown to be greatly elevated in age-related diseases. We demonstrate for the first time that l-DOPA (levodopa) can be biosynthetically incorporated into cell proteins by human cells (THP-1 monocytes and monocyte-derived macrophages). The DOPA-containing proteins generated were selectively visualized on PVDF membranes using a redox-cycling staining method. Many cell proteins contained DOPA and seemed to be synthesized as their full-length forms. The cellular removal of DOPA-containing proteins by THP-1 cells was by proteolysis involving both the proteasomal and the lysosomal systems. The rate of cellular proteolysis of DOPA-containing proteins increased at lower levels of DOPA incorporation but decreased at higher levels of DOPA incorporation. The decreased rate of degradation was accompanied by an increase in the activity of cathepsins B and L but the activity of cathepsin S increased only at lower levels of DOPA incorporation. These data raise the possibility that PB-DOPA could be generated in vivo from l-DOPA, which is the most widely used treatment for Parkinson disease.

AB - Protein-bound 3,4-dihydroxyphenylalanine (PB-DOPA) is a major product of hydroxyl radical attack on tyrosine residues of proteins. Levels of PB-DOPA in cells and tissues have been shown to be greatly elevated in age-related diseases. We demonstrate for the first time that l-DOPA (levodopa) can be biosynthetically incorporated into cell proteins by human cells (THP-1 monocytes and monocyte-derived macrophages). The DOPA-containing proteins generated were selectively visualized on PVDF membranes using a redox-cycling staining method. Many cell proteins contained DOPA and seemed to be synthesized as their full-length forms. The cellular removal of DOPA-containing proteins by THP-1 cells was by proteolysis involving both the proteasomal and the lysosomal systems. The rate of cellular proteolysis of DOPA-containing proteins increased at lower levels of DOPA incorporation but decreased at higher levels of DOPA incorporation. The decreased rate of degradation was accompanied by an increase in the activity of cathepsins B and L but the activity of cathepsin S increased only at lower levels of DOPA incorporation. These data raise the possibility that PB-DOPA could be generated in vivo from l-DOPA, which is the most widely used treatment for Parkinson disease.

KW - DOPA

KW - Proteasome

KW - Lysosome

KW - Cathepsin

KW - Protein oxidation

KW - Free radicals

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