Identification of molting fluid carboxypeptidase A (MF-CPA) in Bombyx mori.
Using microarray analyses, we identified carboxypeptidase A (MF-CPA), which was induced during pupal ecdysis in the wing discs of Bombyx mori. Here, we report the functional characterization of MF-CPA. MF-CPA has amino acid sequence similarities with the proteins in the carboxypeptidase A/B subfamily, from human to nematode. The MF-CPA gene is expressed during the molting periods in the epithelial tissues. MF-CPA is detected in the molting fluid, which fills the space between the old and new cuticle during molting. By Western blot analysis, we show that MF-CPA is secreted as a zymogen and processed in the molting fluid. Recombinant MF-CPA expressed in the insect cells has carboxypeptidase A activity. We propose that MF-CPA degrades the proteins from the old cuticle during the molting periods and contributes to recycling of the amino acids.
Ote
Manabu
M
Department of Agricultural and Environmental Biology, University of Tokyo Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Mita
Kazuei
K
Kawasaki
Hideki
H
Daimon
Takaaki
T
Kobayashi
Masahiko
M
Shimada
Toru
T
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D012831:Bombyx / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D043422:Carboxypeptidases A / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D018507:Gene Expression Regulation, Developmental; D007814:Larva / Q000201:enzymology; D008969:Molecular Sequence Data; D018978:Molting / Q000502:physiology*; D010802:Phylogeny; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D011994:Recombinant Proteins; D043422:Carboxypeptidases A