Melanins in IGR 1 melanoma cells.
Information on the composition of melanins is obtained by analysis both of 4-amino-3-hydroxyphenylalanine (AHP) after hydriodic acid degradation and of pyrrole-2,3,5-tricarboxylic acid (PTCA) after potassium permanganate oxidation. Analysis of thiazole-4,5-dicarboxylic acid (TDCA) and pyrrole-2,3-dicarboxylic acid (PDCA) after permanganate oxidation, provides additional information on the composition, TDCA on pheomelanin residues, and PDCA on indolic residues without carboxy groups. Using model melanins formed from dopa and cysteinyldopa in different proportions, we found the TDCA/(PTCA+PDCA) ratio to yield a reliable estimate of the relative proportions of pheomelanin and eumelanin. The PDCA/PTCA ratio reflects the relationship between indole residues with and without carboxy groups. We have analyzed degradation products from cultures of IGR 1, an extensively studied melanoma cell line. Cell cultures were harvested after 2, 4, and 7 days. Culture media were changed after 2 days in all series, and also after 4 days in one series harvested at 7 days. Cells without medium change had seven times the amount of melanin found in cultures with medium change. The PDCA/PTCA ratio decreased with increasing amounts of melanin. With increased melanization, eumelanin is increased relatively more than pheomelanin. The cell content of 5-S-cysteinyldopa (5-S-CD) was similar in all cultures, while 6-hydroxy-5-methoxyindole-2-carboxylic acid (6H5MICA), a eumelanin precursor metabolite, was found in increased amounts of media of heavily pigmented cultures.
Odh
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Department of Pharmacology, University of Lund, Sweden.
Hindemith
A
A
Carstam
R
R
Paulson
J
J
Rosengren
E
E
Rorsman
H
H
D002853:Chromatography, Liquid; D003548:Cysteinyldopa / Q000032:analysis; D003998:Dicarboxylic Acids / Q000032:analysis; D006801:Humans; D006868:Hydrolysis; D013058:Mass Spectrometry; D008543:Melanins / Q000032:analysis* / Q000737:chemistry / Q000145:classification; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D014442:Monophenol Monooxygenase / Q000032:analysis; D009363:Neoplasm Proteins / Q000032:analysis; D010084:Oxidation-Reduction; D011196:Potassium Permanganate / Q000494:pharmacology; D011758:Pyrroles / Q000032:analysis; D013455:Sulfur / Q000032:analysis; D013844:Thiazoles / Q000032:analysis; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
D003998:Dicarboxylic Acids; D008543:Melanins; D009363:Neoplasm Proteins; D011758:Pyrroles; D013844:Thiazoles; C018362:pheomelanin; D011196:Potassium Permanganate; C093674:pyrrole-2,3-dicarboxylic acid; C041877:eumelanin; D003548:Cysteinyldopa; C083678:thiazole-4,5-dicarboxylic acid; D014443:Tyrosine; D013455:Sulfur; C071227:4-amino-3-hydroxyphenylalanine; C071226:pyrrole-2,3,5-tricarboxylic acid; D014442:Monophenol Monooxygenase