Chitinases of the avian malaria parasite Plasmodium gallinaceum, a class of enzymes necessary for parasite invasion of the mosquito midgut.
The Plasmodium ookinete produces chitinolytic activity that allows the parasite to penetrate the chitin-containing peritrophic matrix surrounding the blood meal in the mosquito midgut. Since the peritrophic matrix is a physical barrier that the parasite must cross to invade the mosquito, and the presence of allosamidin, a chitinase inhibitor, in a blood meal prevents the parasite from invading the midgut epithelium, chitinases (3.2.1.14) are potential targets of malaria parasite transmission-blocking interventions. We have purified a chitinase of the avian malaria parasite Plasmodium gallinaceum and cloned the gene, PgCHT1, encoding it. PgCHT1 encodes catalytic and substrate-binding sites characteristic of family 18 glycohydrolases. Expressed in Escherichia coli strain AD494 (DE3), recombinant PgCHT1 was found to hydrolyze polymeric chitin, native chitin oligosaccharides, and 4-methylumbelliferone derivatives of chitin oligosaccharides. Allosamidin inhibited recombinant PgCHT1 with an IC(50) of 7 microM and differentially inhibited two chromatographically separable P. gallinaceum ookinete-produced chitinase activities with IC(50) values of 7 and 12 microM, respectively. These two chitinase activities also had different pH activity profiles. These data suggest that the P. gallinaceum ookinete uses products of more than one chitinase gene to initiate mosquito midgut invasion.
Vinetz
J M
JM
World Health Organization Collaborating Center for Tropical Diseases, Department of Pathology, the University of Texas Medical Branch, Galveston, Texas 77615, USA. jovinetz@utmb.edu
Valenzuela
J G
JG
Specht
C A
CA
Aravind
L
L
Langer
R C
RC
Ribeiro
J M
JM
Kaslow
D C
DC
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D002688:Chitinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D016384:Consensus Sequence; D009033:Culicidae / Q000469:parasitology*; D004064:Digestive System / Q000469:parasitology; D004847:Epithelial Cells / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007700:Kinetics; D008289:Malaria, Avian; D008969:Molecular Sequence Data; D010964:Plasmodium gallinaceum / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D011994:Recombinant Proteins; D002688:Chitinases