Action of endogenous proteases on the distribution of tyrosinase isozymes in Harding-Passey mouse melanoma.
A high percentage of the total tyrosinase found in Harding-Passey mouse melanoma occurs as a soluble form. This paper shows that melanosomal tyrosinase can be solubilized by several endogenous proteases to yield active tyrosinase. This enzyme, once proteolytically solubilized, can be further degraded, leading to enzyme inactivation. The nature and specificity of the main proteases involved in the solubilization process change depending on the size and necrosis stage of the tumour. Cathepsin B could be the main protease responsible for the solubilization in small tumours (less than 0.5 g). Large tumours are rich in necrotic cells, and cathepsin D and serine-proteases are the main hydrolytic enzymes involved in the proteolytic action on melanosomes. These results support the view that the high activity of tyrosinase found in the soluble fraction of malignant melanoma is mainly an artefact resulting from degradation of melanosomes by a variety of endogenous proteases, rather than the result of the actual occurrence of high levels of an independent cytosolic isozyme.
Martinez
J H
JH
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Murcia, Spain.
Solano
F
F
Lozano
J A
JA
D000818:Animals; D004156:Catechol Oxidase / Q000378:metabolism*; D002402:Cathepsin D / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism*; D007976:Leupeptins / Q000494:pharmacology; D008544:Melanocytes / Q000201:enzymology; D008546:Melanoma, Experimental / Q000201:enzymology*; D051379:Mice; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010436:Pepstatins / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D012995:Solubility
D007527:Isoenzymes; D007976:Leupeptins; D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D010664:Phenylmethylsulfonyl Fluoride; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase; D010447:Peptide Hydrolases; D002402:Cathepsin D; C032854:leupeptin; C031375:pepstatin