Comparison of mono- and dichlorinated tyrosines with carbonyls for detection of hypochlorous acid modified proteins.
Hypochlorous acid is a potent oxidant capable of oxidizing and chlorinating proteins. Based on its indiscriminant reactivity, it is proposed to play a major role in tissue damage associated with a range of inflammatory diseases. We have determined the relative tendencies for formation of protein carbonyls, chlorinated tyrosine residues, and epitopes recognized by an antibody raised against hypochlorous acid oxidized protein (HOP-1) when albumin is treated with hypochlorous acid. We have also tested the specificity of the HOP-1 antibody by measuring how effectively it recognizes proteins oxidized by hypobromous acid. 3-Chlorotyrosine, along with a new marker of hypochlorous acid dependent protein modification, 3, 5-dichlorotyrosine, was formed at the lowest doses of hypochlorous acid that were capable of generating protein carbonyls. Comparatively high doses of hypochlorous acid were needed to generate epitopes recognized by HOP-1, which were also produced by hypobromous acid. Our study demonstrates that it is advantageous to measure protein carbonyls and HOP-1 epitopes in conjunction with chlorinated tyrosines when attempting to identify the oxidants responsible for inflammatory tissue damage.
Chapman
A L
AL
Free Radical Research Group, Christchurch School of Medicine, Christchurch, New Zealand. anna.chapman@chmeds.ac.nz
Senthilmohan
R
R
Winterbourn
C C
CC
Kettle
A J
AJ
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D001959:Bromates / Q000378:metabolism / Q000494:pharmacology; D002255:Carbonic Acid / Q000378:metabolism*; D002417:Cattle; D002713:Chlorine / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism* / Q000494:pharmacology; D007249:Inflammation / Q000378:metabolism; D013058:Mass Spectrometry; D016877:Oxidants / Q000378:metabolism*; D018384:Oxidative Stress; D012709:Serum Albumin / Q000276:immunology / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D000906:Antibodies; D001959:Bromates; D000939:Epitopes; D016877:Oxidants; D012709:Serum Albumin; D014443:Tyrosine; D002255:Carbonic Acid; D002713:Chlorine; D006997:Hypochlorous Acid; C027664:hypobromous acid; C408691:3,5-dichlorotyrosine; C087259:3-chlorotyrosine