title: Gill (Na+,K+)-ATPase in diadromous, freshwater palaemonid shrimps: species-specific kinetic characteristics and alpha-subunit expression.
abstract: To better comprehend physiological adaptation to dilute media and the molecular mechanisms underlying ammonia excretion in palaemonid shrimps, we characterized the (Na+,K+)-ATPase from Macrobrachium amazonicum gills, disclosing high- (K(0.5) = 4.2+/-0.2 micromol L(-1); V = 33.9+/-1.9 U mg(-1)) and low-affinity (K(0.5) = 0.144+/-0.010 mmol L(-1); V = 232.9+/-15.3 U mg(-1)) ATP hydrolyzing sites. Stimulation by Na+ (K(0.5) = 5.5+/-0.3 mmol L(-1); V = 275.1+/-15.1 U mg(-1)), Mg2+ (K(0.5) = 0.79+/-0.06 mmol L(-1); V = 261.9+/-18.3 U mg(-1)), K+ (K(M) = 0.88+/-0.04 mmol L(-1); V = 271.8+/-10.9 U mg(-1)) and NH4(+) (K(M) = 5.0+/-0.2 mmol L(-1); V = 385.9+/-15.8 U mg(-1)) obeys single saturation curves, activity being stimulated synergistically by NH4(+) and K+. There is a single K+ binding site, NH4(+) binding to a second, exclusive site, stimulating activity by 33%, modulating K+ affinity. (Na+,K+)-ATPase activity constitutes approximately 80% of total ATPase activity (K(Iouabain) = 147.5+/-8.9 micromol L(-1)); Na+-, K+-, Ca2+-, V- and F(o)F(1)-ATPases are also present. M. amazonicum microsomal fractions possess approximately 2-fold less (Na+,K+)-ATPase alpha-subunit than M. olfersi, consistent with a 2.6-fold lower specific activity. These differences in (Na+, K+)-ATPase stimulation by ATP and ions, and specific activities of other ATPases, suggest the presence of distinct biochemical adaptations to life in fresh water in these related species.
authors.affiliation: Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-901, SP, Brazil.
authors.forename: L C F
authors.initials: LC
authors.lastname: Santos
authors.forename: N M
authors.initials: NM
authors.lastname: Belli
authors.forename: A
authors.initials: A
authors.lastname: Augusto
authors.forename: D C
authors.initials: DC
authors.lastname: Masui
authors.forename: F A
authors.initials: FA
authors.lastname: Leone
authors.forename: J C
authors.initials: JC
authors.lastname: McNamara
authors.forename: R P M
authors.initials: RP
authors.lastname: Furriel
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003445:Crustacea; D004591:Electrophoresis, Polyacrylamide Gel; D005880:Gills / Q000201:enzymology*; D007700:Kinetics; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D000254:Sodium-Potassium-Exchanging ATPase