title: cDNA cloning of prophenoloxidase from the freshwater crayfish Pacifastacus leniusculus and its activation.
abstract: Prophenoloxidase (proPO), an enzyme that is the terminal component of the so-called proPO activating system, a defense and recognition system in crustaceans and insects, has been purified and cloned from a crayfish blood cell cDNA library. The deduced amino acid sequence codes for a polypeptide with a mass of 80,732 Da, which is close to 76 kDa, the apparent mass of the purified enzyme. proPO contains two copper atoms, and two putative copper-binding sites were found in the deduced amino acid sequence. Sequence comparisons show that these putative copper-binding sites are similar to the corresponding sites in arthropod hemocyanins and also, although the sequence similarities are less extensive, similar to tyrosinases from vertebrates and microorganisms. The purified enzyme is a typical tyrosinase because it hydroxylates monophenols and oxidizes o-diphenols but does not oxidize p-diphenols. If a homogeneous preparation of crayfish proPO were incubated with a homogeneous sample of the proPO activating enzyme, a serine proteinase, the cleavage of proPO by this trypsin-like enzyme was found to occur between Arg-176 and Thr-177.
authors.affiliation: Department of Physiological Botany, University of Uppsala, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Aspán
authors.forename: T S
authors.initials: TS
authors.lastname: Huang
authors.forename: L
authors.initials: L
authors.lastname: Cerenius
authors.forename: K
authors.initials: K
authors.lastname: Söderhäll
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003400:Astacoidea / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D004156:Catechol Oxidase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D004792:Enzyme Precursors; C038451:pro-phenoloxidase; D004156:Catechol Oxidase