title: Characterization of an ecto-phosphatase activity in malpighian tubules of hematophagous bug Rhodnius prolixus.
abstract: We have characterized a phosphatase activity present on the external surface of intact Malpighian tubules in Rhodnius prolixus. This phosphatase hydrolyses the substrate p-nitrophenyl phosphate at a rate of 3.38 +/- 0.07 nmol Pi x mg(-1) x min(-1). Phosphatase activity decreased with the increase of the pH from 6.4 to 7.6 pH, a range in which tubules cellular integrity was maintained for at least 1 h. Classical inhibitors of acid phosphatase, such as ammonium molybdate, fluoride, vanadate, mpV-PIC, and bpV-PHEN, caused different patters of inhibition. The ecto-phosphatase present an apparent Km of 1.67 +/- 0.34 mM and Vmax of 5.71 +/- 0.37 nmol Pi x mg(-1) x min(-1) for p-NPP. Zinc chloride inhibited 78.2% of ecto-phosphatase activity, with Ki of 0.35 mM. Such inhibition was reversed by incubation with cysteine and GSH, but not DTT, serine, and GSSG, showing that cysteine residues are important for enzymatic activity. Phosphatase activity increased 141% three days after blood meal, and returned to basal levels 2 days later. These results suggest that ecto-phosphatase activity could be involved in a diuretic mechanism, essential in the initial days after a blood meal for the control of Rhodnius homeostasis.
authors.affiliation: Departamento de Bioquímica Médica, ICB, Universidade Federal do Rio de Janeiro, UFRJ, Cidade Universitária, Ilha do Fundão, 21941-590, Rio de Janeiro, RJ, Brazil.
authors.forename: Rodrigo B
authors.initials: RB
authors.lastname: Ferraro
authors.forename: Joana L
authors.initials: JL
authors.lastname: Sousa
authors.forename: Renata D C
authors.initials: RD
authors.lastname: Cunha
authors.forename: José R
authors.initials: JR
authors.lastname: Meyer-Fernandes
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008317:Malpighian Tubules / Q000166:cytology / Q000201:enzymology*; D009752:Nutritional Status / Q000502:physiology*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D012239:Rhodnius / Q000201:enzymology*; D013997:Time Factors
chemical_list: D010744:Phosphoric Monoester Hydrolases