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Characterization of Multicopper Oxidase Related Protein (MCORP) in Two Insect Species
Author(s) -
PENG ZEYU,
Gorman Maureen J,
Arakane Yasuyuki,
Kanost Michael R
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.854.6
Subject(s) - biology , anopheles gambiae , bombyx mori , gene , multicopper oxidase , rna interference , genetics , drosophila melanogaster , recombinant dna , insect , biochemistry , enzyme , botany , rna , laccase , malaria , immunology
Multicopper oxidases (MCOs) have diverse functions but are evolutionarily and structurally related. Ten histidines and one cysteine form two highly conserved copper binding centers, which are required for oxidase activity. We discovered a gene that we named multicopper oxidase related protein (MCORP), which looks somewhat like a MCO, but lacks the conserved cysteine and many of the histidines. We identified MCORP orthologs in many insect species, including Tribolium castaneum, Anopheles gambiae, Aedes aegypti, Culex pipiens, Bombyx mori, and Acyrthosiphon pisum . Phylogenetic analysis of MCORPs and MCOs indicated that MCORPs belong to a specific cluster. We cloned full‐length cDNAs of A. gambiae and T. castaneum MCORP and studied their expression patterns by RT‐PCR. Both genes are highly expressed in late developmental stages, particularly in pupae. In adults, AgMCORP is expressed predominantly in reproductive tissues. We purified recombinant AgMCORP and TcMCORP to make antibodies that we will use to determine the location of MCORP within an insect. RNAi mediated knock down of MCORP in T. castaneum resulted in a pupal lethal phenotype, which demonstrated that TcMCORP is essential to pupal development. To try to learn the function of MCORP, we will expand our RNAi studies and determine whether recombinant MCORPs have enzymatic activity. This research was supported by a grant from NIH (R01 AI070864).