Isolation of nitrotyrosine-containing peptides by using an insoluble-antibody column.
Antibodies to nitrotyrosine were prepared in goats or rabbits by injecting a nitrotyrosine-protein conjugate. The antibodies were purified by using a protein that had been nitrated with tetranitromethane. These antibodies were used to isolate nitrotyrosine-containing peptides from nitrated lysozyme. The nitrotyrosine-containing peptides were thus purified (55% yield) in one step and the positions of nitration in lysozyme were found to be at tyrosine-20 and tyrosine-23. This method is of general applicability for the determination of the position of nitrotyrosine in proteins.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D002417:Cattle; D002846:Chromatography, Affinity; D004586:Electrophoresis; D006041:Goats / Q000276:immunology; D006241:Haptens; D007114:Immunization; D008722:Methods; D009113:Muramidase / Q000032:analysis; D009574:Nitro Compounds / Q000302:isolation & purification; D010455:Peptides / Q000302:isolation & purification*; D011817:Rabbits / Q000276:immunology; D012710:Serum Albumin, Bovine; D014443:Tyrosine / Q000302:isolation & purification*
D000596:Amino Acids; D000906:Antibodies; D006241:Haptens; D009574:Nitro Compounds; D010455:Peptides; D012710:Serum Albumin, Bovine; D014443:Tyrosine; D009113:Muramidase