Cooperation of dopachrome conversion factor with phenoloxidase in the eumelanin pathway in haemolymph of Locusta migratoria (Insecta).
Dopachrome Conversion Factor (DCF) was found in the plasma of the locust Locusta migratoria. It has an apparent molecular mass of 85,000. Its K(m) was 0.2 mM at 22 degrees C and pH 7 with L-dopachrome as substrate. It had a high substrate specificity for L-dopachrome, methyl-L-dopachrome and L-dopachrome methyl ester but no activity on the corresponding D-isomers or on dopaminechrome. DCF was devoid of any phenoloxidase activity. Under action of DCF, L-dopachrome was converted into dihydroxyindole, which showed that a decarboxylation occured in the course of reaction. Locust DCF was inhibited by indole-3-propionic acid but not by indole-3- or indole-2-carboxylic acid. It was also inhibited by L-tryptophan in a competitive manner. Inhibition and substrate specificity suggest that a carboxyl group, either free or as a methyl ester, is necessary but not sufficient for enzyme recognition. When purified prophenoloxidase was activated and then added to dihydroxyindole either prepared by chemical synthesis or obtained by action of purified DCF on dopachrome, black pigments with a maximum absorption at 540 nm were generated. Therefore in the eumelanin pathway of locust plasma, phenoloxidase can catalyze the reaction that converts the product generated by DCF.
Cherqui
A
A
Laboratoire de Pathologie Comparée, INRA-CNRS (URA 2209), Montpellier, France.
Duvic
B
B
Reibel
C
C
Brehélin
M
M
D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D006110:Grasshoppers / Q000201:enzymology*; D006458:Hemolymph / Q000201:enzymology*; D045563:Indolequinones; D007211:Indoles / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D008543:Melanins / Q000378:metabolism*; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D011809:Quinones / Q000378:metabolism; D013379:Substrate Specificity; D014443:Tyrosine / Q000378:metabolism
D045563:Indolequinones; D007211:Indoles; D008543:Melanins; D011809:Quinones; C041877:eumelanin; C001123:dopachrome; D014443:Tyrosine; D014442:Monophenol Monooxygenase; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase; C033871:5,6-dihydroxyindole