Characterization of (Na+, K+)-ATPase in gill microsomes of the freshwater shrimp Macrobrachium olfersii

Furriel, Rosa dos Prazeres Melo; McNamara, John Campbell; Leone, Francisco de Assis

doi:10.1016/s0305-0491(00)00184-x

Cited by 91 publications

(87 citation statements)

References 41 publications

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“…The species thus represents an attractive model in which to examine the role of the (Na þ , K þ )-ATPase in gill ion transport. Recent results from our laboratory (Furriel et al, 2000) reveal that the kinetic characteristics of the enzyme corroborate a model proposed for Na þ uptake across the gill epithelium of M. olfersii (McNamara and Torres,'99). Na þ -and K þ -activated ATP hydrolysis is a widely distributed, phylogenetically conserved key mechanism, which has lead to extensive investigation of (Na þ , K þ )-ATPase structure-function relationships, particularly in vertebrates (Kaplan, 2002, Scheiner-Bobis, 2002.…”

supporting

confidence: 57%

“…4), in the presence or absence of 50 mmol.L À1 NH 4 þ ions, reveals that the NH 4 þ ions directly affect (Na þ ,K þ )-ATPase activity, despite other ATPases present in the microsomal preparation (Furriel et al, 2000). Like the enzyme from C. danae gill tissue (Masui et al, 2002), the K I in the presence of NH 4 þ ions was roughly 67% less than in their absence.…”

Section: Nhmentioning

confidence: 97%

“…Maximum velocity (V), the apparent dissociation constant (K 0.5 ), the Michaelis-Menten constant (K M ), the Hill coefficient (n H ) and the enzyme-inhibitor complex dissociation constant (K I ) were calculated as described by Furriel et al (2000). The kinetic parameters given in the Tables are the mean7SD and were calculated from at least three (NZ3) different gill microsomal preparations.…”

Section: Estimation Of Kinetic Parametersmentioning

confidence: 99%

“…Total ATPase activity was routinely assayed at 251C using a GAPDH/PGK linked system in which the hydrolysis of ATP was coupled to the reduction of NAD þ (Furriel et al, 2000). The reduction of NAD þ was monitored at 340 nm (E 340 nm, pH 7.5 ¼ 6200 M À1 .cm À1 ) in a Hitachi U-3000 spectrophotometer equipped with thermostatted cell holders.…”

Section: Measurement Of Atpase Activitymentioning

confidence: 99%

“…Na þ -and K þ -activated ATP hydrolysis is a widely distributed, phylogenetically conserved key mechanism, which has lead to extensive investigation of (Na þ , K þ )-ATPase structure-function relationships, particularly in vertebrates (Kaplan, 2002, Scheiner-Bobis, 2002. The (Na þ , K þ )-ATPase (Mo and Greenaway, 2001;Furriel et al, 2000Furriel et al, , 2001Wilder et al, 2000;Mananes et al, 2002;Masui et al, 2002Masui et al, , 2003 and K þ -and Na þ -stimulated ATPases of crustacean gills have also been widely studied (Moretti et al,'91;Proverbio et al,'91;Lima et al,'97). The energy of ATP hydrolysis drives the coupled extrusion of Na þ and uptake of K þ ions by the (Na þ ,K þ )-ATPase across the plasma membrane of most eukaryotic cells.…”

mentioning

confidence: 99%

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Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

et al. 2003

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The effect of NH 4 þ ions on (Na þ ,K þ )-ATPase hydrolytic activity was examined in a gill microsomal fraction from M. olfersii. In the absence of NH 4 þ ions, K þ ions stimulated ATP hydrolysis, exhibiting cooperative kinetics (n H ¼0.8), to a maximal specific activity of V¼556. þ and K þ ions. These data suggest that at high NH 4 þ ion concentrations, the (Na þ , K þ )-ATPase exposes a new site; the subsequent binding of NH 4 þ ions stimulates ATP hydrolysis to rates higher than those for K þ ions alone. This is the first demonstration that (Na þ , K þ )-ATPase activity in a freshwater shrimp gill is modulated by ammonium ions, independently of K þ ions, an effect that may constitute a fine-tuning mechanism of physiological relevance to osmoregulatory and excretory processes in palaemonid shrimps.

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supporting

confidence: 57%

Section: Nhmentioning

confidence: 97%

Section: Estimation Of Kinetic Parametersmentioning

confidence: 99%

Section: Measurement Of Atpase Activitymentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

et al. 2003

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Low salinity‐induced alterations in epithelial ultrastructure, Na⁺/K⁺‐ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae)

et al. 2017

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Fresh caught Clibanarius vittatus [SW, 31‰ salinity (S)] were acclimated to a dilute medium (15‰ S) for 10 days, employing silver staining to locate gill ion transporting tissue, immunofluorescence to localize the Na/K-ATPase α-subunit in the lamellae, and electron microscopy to portray ultrastructural changes in the gill epithelia. Na/K-ATPase activity was characterized kinetically in a gill microsomal fraction, including synergistic stimulation by NH plus K. Silver staining revealed that all 26 phyllobranchiate arthro- and pleurobranchiae participate in ion transport. Na/K-ATPase α-subunit staining was weak in SW crabs and distributed exclusively and irregularly within the intralamellar septal cells, particularly at the septal-pillar cell body junctions, and septal cell cytoplasm facing the hemolymph space. In 15‰ S crabs, α-subunit localization was intense, occupying the entire thickened septum. Pillar cells and flanges did not stain. Mitochondria and membrane foldings increased in the pillar cell flanges and intralamellar septal cells, greatly amplifying surface area. Only a single ATP binding site (V = 130.8 ± 10.5 nmol min mg protein; K = 55.3 ± 1.7 μmol l) obeying Michaelis-Menten kinetics was disclosed. Na/K-ATPase activity was modulated by Mg, Na, and NH, exhibiting site-site interactions; K modulation showed Michaelis-Menten kinetics. K plus NH synergistically stimulated activity ≈ 1.7-fold. Ouabain inhibited total ATPase activity by ≈ 70% (K = 220-300 μmol l), revealing phosphohydrolytic activities other than the Na/K-ATPase. Despite ample phylogenetic separation, the phyllobranchiate lamellae of the Anomura and Caridea share many ultrastructural features, that is, an intralamellar septum and opposed abutting pillar cells, similar Na/K-ATPase distribution, and comparable kinetic characteristics. These findings suggest either convergent evolution at the structural and biochemical levels, or preservation of traits present in a remote common ancestor.

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Structural and biochemical correlates of Na⁺,K⁺‐ATPase driven ion uptake across the posterior gill epithelium of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)

et al. 2010

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To better comprehend the structural and biochemical underpinnings of ion uptake across the gills of true freshwater crabs, we performed an ultrastructural, ultracytochemical and morphometric investigation, and kinetically characterized the Na(+),K(+)-ATPase, in posterior gill lamellae of Dilocarcinus pagei. Ultrastructurally, the lamellar epithelia are markedly asymmetrical: the thick, mushroom-shaped, proximal ionocytes contain elongate mitochondria (41% cell volume) associated with numerous (≈14 µm² membrane per µm³cytoplasm), deep invaginations that house the Na(+),K(+)-ATPase, revealed ultracytochemically. Their apical surface is amplified (7.5 µm² µm⁻²)) by stubby evaginations whose bases adjoin mitochondria below the subcuticular space. The apical membrane of the thin, distal ionocytes shows few evaginations (1.6 µm² µm⁻²), each surrounding a mitochondrion, abundant in the cytoplasm below the subcuticular space; basolateral invaginations and mitochondria are few. Fine basal cytoplasmic bridges project across the hemolymph space, penetrating into the thick ionocytes, suggesting ion movement between the epithelia. Microsomal Na(+),K(+)-ATPase specific activity resembles marine crabs but is ≈5-fold less than in species from fluctuating salinities, and freshwater shrimps, suggesting ion loss compensation by strategies other than Na(+) uptake. Enzyme apparent K(+) affinity attains 14-fold that of marine crabs, emphasizing the relevance of elevated K(+) affinity to the conquest of fresh water. Western blotting and biphasic ouabain inhibition disclose two α-subunit isoforms comprising distinct functional isoenzymes. While enzyme activity is not synergistically stimulated by NH(4) (+) and K(+), each increases affinity for the other, possibly assuring appropriate intracellular K(+) concentrations. These findings reveal specific structural and biochemical adaptations that may have allowed the establishment of the Brachyura in fresh water.

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Characterization of (Na+, K+)-ATPase in gill microsomes of the freshwater shrimp Macrobrachium olfersii

Cited by 91 publications

References 41 publications

Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Low salinity‐induced alterations in epithelial ultrastructure, Na⁺/K⁺‐ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae)

Structural and biochemical correlates of Na⁺,K⁺‐ATPase driven ion uptake across the posterior gill epithelium of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)

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Characterization of (Na+, K+)-ATPase in gill microsomes of the freshwater shrimp Macrobrachium olfersii

Cited by 91 publications

References 41 publications

Modulation of gill Na+,K+‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Modulation of gill Na+,K+‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Low salinity‐induced alterations in epithelial ultrastructure, Na+/K+‐ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae)

Structural and biochemical correlates of Na+,K+‐ATPase driven ion uptake across the posterior gill epithelium of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)

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Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Modulation of gill Na⁺,K⁺‐ATPase activity by ammonium ions: Putative coupling of nitrogen excretion and ion uptake in the freshwater shrimp Macrobrachium olfersii

Low salinity‐induced alterations in epithelial ultrastructure, Na⁺/K⁺‐ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae)

Structural and biochemical correlates of Na⁺,K⁺‐ATPase driven ion uptake across the posterior gill epithelium of the true freshwater crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)