title: Kinetic properties of catecholoxidase activity of tarantula hemocyanin.
abstract: Phenoloxidases occur in almost all organisms, being essentially involved in various processes such as the immune response, wound healing, pigmentation and sclerotization in arthropods. Many hemocyanins are also capable of phenoloxidase activity after activation. Notably, in chelicerates, a phenoloxidase has not been identified in the hemolymph, and thus hemocyanin is assumed to be the physiological phenoloxidase in these animals. Although phenoloxidase activity has been shown for hemocyanin from several chelicerate species, a characterization of the enzymatic properties is still lacking. In this article, the enzymatic properties of activated hemocyanin from the tarantula Eurypelma californicum are reported, which was activated by SDS at concentrations above the critical micellar concentration. The activated state of Eurypelma hemocyanin is stable for several hours. Dopamine is a preferred substrate of activated hemocyanin. For dopamine, a K(M) value of 1.45 +/- 0.16 mm and strong substrate inhibition at high substrate concentrations were observed. Typical inhibitors of catecholoxidase, such as l-mimosine, kojic acid, tyramine, phenylthiourea and azide, also inhibit the phenoloxidase activity of activated hemocyanin. This indicates that the activated hemocyanin behaves as a normal phenoloxidase.
authors.affiliation: Institut für Molekulare Biophysik, Johannes Gutenberg Universität, Mainz, Germany.
authors.forename: Elmar
authors.initials: E
authors.lastname: Jaenicke
authors.affiliation: Institut für Molekulare Biophysik, Johannes Gutenberg Universität, Mainz, Germany.
authors.forename: Heinz
authors.initials: H
authors.lastname: Decker
mesh_terms: D000818:Animals; D004156:Catechol Oxidase / Q000737:chemistry* / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006433:Hemocyanins / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013112:Spiders / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D012967:Sodium Dodecyl Sulfate; D006433:Hemocyanins; D004156:Catechol Oxidase