cpbAg1 encodes an active carboxypeptidase B expressed in the midgut of Anopheles gambiae.
We previously used differential display to identify several Anopheles gambiae genes, whose expression in the mosquito midgut was regulated upon ingestion of Plasmodium falciparum. Here, we report the characterization of one of these genes, cpbAg1, which codes for the first zinc-carboxypeptidase B identified in An. gambiae and in any insect. Expression of cpbAg1 in baculovirus gave rise to an active enzyme, and determination of the N-terminal amino acids confirmed that CPBAg1 contains a signal peptide and a pro-peptide, typical features of digestive zinc carboxypeptidases. cpbAg1 mRNA was mainly produced in the mosquito midgut, where it accumulated in unfed females and was rapidly down-regulated upon blood feeding. Annotation of the An. gambiae genome predicts twenty-three sequences coding for zinc-carboxypeptidases of which only two (cpbAg1 and cpbAg2) are expressed at a significant level in the mosquito midgut.
Lavazec
C
C
Unité de Biologie et Génétique du Paludisme, Institut Pasteur, Paris, France.
Bonnet
S
S
Thiery
I
I
Boisson
B
B
Bourgouin
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D043424:Carboxypeptidase B / Q000235:genetics* / Q000378:metabolism; D016000:Cluster Analysis; D017124:Conserved Sequence / Q000235:genetics; D017931:DNA Primers; D015536:Down-Regulation / Q000502:physiology*; D041981:Gastrointestinal Tract / Q000378:metabolism; D017344:Genes, Insect / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D010802:Phylogeny; D011487:Protein Conformation; D012333:RNA, Messenger / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D043424:Carboxypeptidase B