Presence of chitinase and beta-N-acetylglucosaminidase in the Aedes aegypti. a chitinolytic system involving peritrophic matrix formation and degradation.
Measurement of the hydrolysis of specific fluorogenic substrates by spectrophotometry as well as the substrate activity-SDS-PAGE gel analysis of the chitinolytic activity in Aedes aegypti guts showed that both chitinase and beta-N-acetylglucosaminidase are present and physiologically active. Both enzymes were present even in guts from unfed insects, but the activities increased rapidly after feeding on blood or an artificial protein-free diet. Chitinase activity was predominantly of the 'endo'-type, reaching its maximum activity at 36 h and then declining to very low levels after the degradation of the peritrophic matrix (PM). Chitinase assay in gels after SDS-PAGE was a very sensitive method that allowed us to detect two chitinases with distinct molecular weights in the mosquito gut. Hydrolysis of a chitinase-specific substrate by chitinolytic activities in the mosquito guts was inhibited by allosamidin, a potent chitinase inhibitor. Allosamidin treatment led to the formation of an atypical thick PM, while the addition of exogenous chitinase completely blocked its formation. This chitinolytic system appears to operate both on the formation and degradation of the PM. Since the PM is involved in pathogen invasion, these results are important in facilitating a search for mechanisms that can block pathogen development in the mosquito vector.
Filho
Benedito P D
BP
Laboratório de Microbiologia, Departamento de Análises Clínicas, Universidade Estadual de Maringá, Av. Colombo, 5790, CEP 87030-121, Maringá, PR, Brazil.
Lemos
Francisco J A
FJ
Secundino
Nágila F C
NF
Páscoa
Valéria
V
Pereira
Sheila T
ST
Pimenta
Paulo F P
PF
D000118:Acetylglucosaminidase / Q000378:metabolism*; D000330:Aedes / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005247:Feeding Behavior; D005260:Female; D007700:Kinetics; D051379:Mice; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D002686:Chitin; D002688:Chitinases; D000118:Acetylglucosaminidase