A gut-specific serine protease from the malaria vector Aanopheles gambiae is downregulated after blood ingestion.
A chymotrypsin-like serine protease gene (AgChyL) was cloned from the mosquito Anopheles gambiae by a polymerase chain reaction (PCR)-based subtractive cDNA cloning strategy. AgChyL messenger RNA (mRNA) is abundant in the adult female gut prior to, and for 8 h following, a blood meal. During the peak of digestion, from 12 to 24 h following a blood meal, AgChyL mRNA abundance decreased to barely detectable levels. AgChyL mRNA was abundant again by 48 h following a blood meal. Recombinant pro-AgChyL was expressed in Escherichia coli. The pro-enzyme can be activated by trypsin. Activated AgChyL cleaves the synthetic chymotrypsin substrate succinyl-L-Ala-Ala-Pro-Phe-nitroanilide, but not two other synthetic chymotrypsin substrates or synthetic trypsin and elastase substrates. The potential role of AgChyL in the coordination of An. gambiae digestion is discussed.
Shen
Z
Z
Case Western Reserve University, School of Medicine Department of Genetics, Cleveland, OH 44106-4955, USA.
Edwards
M J
MJ
Jacobs-Lorena
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D001769:Blood; D002918:Chymotrypsin / Q000145:classification / Q000235:genetics*; D018076:DNA, Complementary; D015536:Down-Regulation; D005247:Feeding Behavior; D005260:Female; D015870:Gene Expression; D017344:Genes, Insect; D019476:Insect Proteins; D007303:Insect Vectors / Q000201:enzymology* / Q000235:genetics; D008288:Malaria; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000145:classification / Q000235:genetics*
D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C412871:ChyL protease, Anopheles; D012697:Serine Endopeptidases; D002918:Chymotrypsin