A highly sensitive method for the determination of protein bound 3,4-dihydroxyphenylalanine as a marker for post-translational protein hydroxylation in human tissues ex vivo.
A highly sensitive, specific and tissue-independent method is described to evaluate oxidative stress-mediated protein hydroxylation in red blood cells, frontal cortex, and liver by HPLC separation and electrochemical detection of protein-bound 3,4-dihydroxyphenylalanine (DOPA) following gas-phase amino acid hydrolysis of tissue protein extracts containing exclusively proteins larger than 3 kDa. Simultaneous measurement of protein tyrosine (Tyr) content using fluorescence detection results in a tissue specific DOPA/Tyr ratio that may reflect oxidative stress-mediated protein modifications in disease, or following the exposure to oxidative stress-inducing agents.
Harth
R
R
University Clinic and Policlinic for Psychiatry and Psychotherapy, Clinical Neurochemistry, D-97080 Würzburg, Germany.
Gerlach
M
M
Riederer
P
P
Götz
M E
ME
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004295:Dihydroxyphenylalanine / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D004563:Electrochemistry / Q000379:methods*; D004912:Erythrocytes / Q000378:metabolism; D005625:Frontal Lobe / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D006900:Hydroxylation; D008099:Liver / Q000378:metabolism; D008970:Molecular Weight; D018384:Oxidative Stress; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014443:Tyrosine / Q000032:analysis / Q000378:metabolism
D011506:Proteins; D014443:Tyrosine; D004295:Dihydroxyphenylalanine