The ARR4 gene from Saccharomyces cerevisiae was highly homological with E. coli arsA. The defective nucleotide-binding domain of ARR4[G30R] exhibited negative dominance from the growth curve experiment, showing that ARR4 protein might act as a dimer in vivo. On the nonreducing gel, the purified 40 kDa ARR4 presented as monomer, dimer and oligomer conformational states. Besides, the chemical cross-linking experiments confirmed the positional size of dimeric ARR4. The metal ions of Co2+, Sb3+, As3+ and As5+ have not effect on the ATPase activity and ARR4 oligomerization. ARR4 exhibit a low level of Mg2+-ATPase activity in the range of 50-70 and 100-120 nmol/min/mg respectively as estimated by phosphate released and NADH-coupled assay. The maximal activity was obtained at a ratio of ATP:Mg2+ of 2:1 and the ATPase activity of ARR4[G30R] was about 50% of that of the ARR4 from both ATPase assays. In the presence of ATP or Mg2+, ARR4 has similar ratios of dimer and monomer. When incubated with Mg2+-ATP, ARR4 tended to form more dimer than with ATP and Mg2+ individually. ARR4[G30R] exhibited less dimer formation. Mixing ARR4 and ARR4[G30R] at a constant total protein concentration with different molar ratios (1:1, 1:2 and 1:4), a near linear relationship of activity versus amount of the ARR4 protein was observed. This observation suggests that ATP hydrolysis takes places in one ATP binding site independent of the other site of NBD. The proportional ATPase activities were consistent with the ratios of ARR4-ARR4[G30R] dimerization on the nonreducing gels. In conclusion, the conformational changes induced by Mg2+-ATP is related to the dimerization of ARR4 and both its NBDs would hydrolyze ATP independently.

Ce travail de thèse vise à améliorer notre compréhension des effets l'entraînement en résistance sur la performance et le muscle strié squelettique. La dynamique de ces effets de l'entraînement a été appréhendée de façon systématique grâce à des outils issus de la théorie des systèmes, auprès de 26 rongeurs entraînés en résistance dans un protocole d'escalade avec charges additionnelles. Le modèle classique (Banister et coll, 1975) a permis de décrire les variations de performance de manière significative (R2 = 0,53, P<0,001). L'origine des gains de performance très marqués (+136% par rapport au groupe contrôle) a été recherchée parmi les mécanismes adaptatifs musculaires potentiels. A l'issue de l'entraînement, une augmentation de l'activité de la myosine ATPase de 123 ± 61% indépendante du phénotype a été observée par rapport aux animaux contrôles. Cette augmentation de la puissance chimique consommée semble liée à une augmentation de la vitesse des étapes d'hydrolyse de l'ATP et surtout de celle de la libération des produits de cette hydrolyse (i.e. ADP et Pi) accompagnant la bascule de la tête de myosine. Une nouvelle forme de plasticité musculaire semble avoir été identifiée. Sur la base des mécanismes adaptatifs musculaires, une nouvelle formulation mathématique plus physiologique du modèle des effets de l'entraînement a été proposée et a aboutit à une meilleure qualité d'ajustement (R2 = 0,71, P<0,001). La fonction impulsionnelle du modèle classique a été remplacée par une fonction exponentielle de croissance qui semble plus appropriée pour rendre compte à la fois des variations de performance mais aussi des adaptations qui surviennent au sein du tissu musculaire comme au sein des unités contractiles elles-mêmes. / This thesis work aims to improve our understanding of the effects of resistance training on performance and skeletal muscle. The dynamic of these effects of training has been apprehended systematically trough tools from systems theory, with 26 rodents resistance trained on a climbing protocol with additional weights. The classical model (Banister et al, 1975) was suitable to analyze the training response (R2 = 0.53, P <0.001). The origin of the very marked performance gains (+ 136% compared to the control group) was investigated among the potential muscle adaptive mechanisms. At the end of the training program, an increase of 123 ± 61% in myosin ATPase activity independent of the phenotype was observed compared to control animals. This increase in myosin ATPase activity seems to occur precisely during the main myosin head isomerization step (i.e. powerstroke) that includes the liberation of the hydrolysis products, and to a lesser extent, during ATP hydrolysis step. A new form of muscular plasticity seems identified. Based on muscle adaptive mechanisms, a new mathematical formulation, more physiological, of the model of the training effects has been proposed and resulted in a better fit (R2 = 0.71, P <0.001). The impulse function of the traditional model has been replaced by an exponential growth function that seems more suitable to analyze both the training response and the adaptations that occur within the muscle tissue as in the contractile units themselves.

Mosquitoes affect human health worldwide as a result of their ability to vector multiple diseases. Mosquitocide resistance is a serious public health challenge that warrants the development of improved chemical control strategies for mosquitoes. Previous studies demonstrate the mosquito blood-brain barrier (BBB) to interfere with the target-site delivery and action of anticholinesterase chemistries. The ATP-binding cassette (ABC) transporters are efflux proteins that assist in maintaining the BBB interface and serve as a first line of defense to mosquitocide exposures. To date, there are three subfamilies (ABC -B, -C, -G) of ABC transporters; however, knowledge of these chemistries interacting with mosquito ABC transporter(s) is limited. Here, I report that tacrine and bis(7)-tacrine are relative non-toxic anticholinesterases at solubility limits; however, the addition of verapamil enhances toxicity of both tacrine and bis(7)-tacrine to mosquitoes. Verapamil significantly increases the mortality of mosquitoes exposed to tacrine and bis(7)-tacrine compared to the tacrine- and bis(7)- tacrine-only treatments. Tacrine and bis(7)-tacrine reduce acetylcholinesterase activity in mosquito head preparations compared to the untreated mosquitoes; however, the addition of verapamil significantly increases the anticholinesterase activity of tacrine and bis(7)-tacrine compared to the tacrine-and bis(7)-tacrine-only treatments. Tacrine and bis(7)-tacrine increase ATPase activity in Aedes aegypti at lower concentrations compared to that of verapamil (Fig. 3). The differential increase in ATPase activity suggests that tacrine and bis(7)-tacrine are more suitable substrates for ABC transporter(s) compared to verapamil and, thus, provides putative evidence that ABC transporter(s) is a pharmacological obstacle to the delivery of these anticholinesterases to their intended target site. / Master of Science in Life Sciences

A Na,K-ATPase é uma proteína encontrada na membrana plasmática de praticamente todas as células animais, que utiliza a energia derivada da hidrólise do ATP para transportar 3 íons Na+ e 2 íons K+. É composta por duas subunidades denominadas e . Um aspecto que ainda gera controvérsias se refere à sua forma de associação nativa e funcional como um protômero ou ainda na forma de oligômeros ()2 ou ()4. Uma forma de estudar essa enzima é pela sua solubilização da membrana, e posteriormente reconstituição em lipossomos de DPPC:DPPE. A caracterização cinética e estrutural desse sistema mostra que a enzima se apresenta na forma oligomérica ()2. O objetivo desse trabalho foi avaliar os mecanismos de dissociação e de desnaturação da Na,K-ATPase solubilizada bem como da reconstituída em lipossomos de DPPC:DPPE, por agentes físicos (temperatura) e químicos (relação proteína:detergente, uso de agentes caotrópicos como a guanidina e mudanças de pH), para interpretar as suas formas de associação e regulação. Para isso, foram realizados experimentos de dicroísmo circular (CD), calorimetria (DSC), infravermelho (FTIR), fluorescência de emissão do triptofano, tensão superficial, elasticidade, atividade catalítica (ATPase e pNPPase). Os estudos de CD em função da variação de temperatura mostraram que ocorre uma transição na curva de elipticidade (222 nm) a 43,7°C para a enzima solubilizada e a 42,0°C para a enzima reconstituída em lipossomos. Estas transições foram também encontradas pela técnica de FTIR. Os experimentos por DSC para a enzima solubilizada revelaram a presença de três picos em 54,7; 64,7 e 67,8°C. Já para a enzima reconstituída observam-se transições em menores temperaturas entre 30 a 40ºC (referentes aos lipídios) e ainda a preservação do pico de transição para proteína em 68,0°C. A análise de fluorescência de triptofano para ambas formas de enzima revelou deslocamentos de pico máximo de emissão a partir de 60°C. Já a presença de guanidina mostrou dois pontos de transição em 3 e 5 mol.L-1 para a Na,K-ATPase solubilizada. O efeito de diferentes meios tamponantes revelou que a enzima apresenta maiores conteúdos em -hélice em pH 7,5, concomitante com um aumento na intensidade de emissão de fluorescência do triptofano na faixa de pH de 5,0 a 8,5. Analisando conjuntamente todas as técnicas podemos propor um mecanismo de dissociação/desnaturação da enzima em função da temperatura. Primeiramente a enzima passa do seu estado oligomérico ()2 e forma protômeros . A atividade ATPase é perdida completamente (acima de 60ºC) quando as subunidades são completamente separadas, ocorrendo então uma agregação das subunidades , através dos domínios citoplasmáticos. Finalmente, a análise da enzima em diferentes proporções de proteína:detergente revela que a Na,K-ATPase, na presença de concentrações abaixo da CMC, se encontra na forma ()2 ou ainda ()4 (dependendo da concentração de proteína). Já para concentrações acima da CMC ocorre a separação das subunidades e consequente perda de atividade catalítica. Devido à dependência da atividade ATPase com seu estado conformacional e seu estado de oligomerização, este estudo realizado por técnicas bioquímicas e biofísicas, resulta em novas informações acerca da compreensão dos mecanismos que controlam o processo de associação, o qual é importante para a função da enzima na membrana natural. / Na,K-ATPase is a protein found in the plasmatic membrane of almost all animal cells and it uses the energy from ATP hydrolysis to transport 3 Na+ ions and 2 K+ ions. It is formed by subunits called and. One controversial aspect refers to its native and functional association form as a protomer or still in ()2 or ()4 oligomers form. One way to study this protein in our laboratory is by its solubilization from membrane, and later reconstitution in liposome from DPPC:DPPE. The kinetic and structural characterization fo this system shows that the enzyme presents itself in the oligomeric form ()2. The aim of this work was to evaluate the dissociation and denaturation mechanisms of the solubilized NA,K-ATPase as well as the one reconstituted in DPPC:DPPE liposome, by physic (temperature) and chemical agents (relation protein:detergent, use of chaotropic agents as Guanidine chloride, or still by the pH changes, to interpret its association and regulation forms. To that end, experiments of circular dichroism (CD), calorimetry (DSC), superficial tension, elasticity , catalytic activity (ATPase an pNPPase) were done. The CD studies in function of temperature variation have shown that a transition occurs in the ellipticity curve (222 nm) at 43.7ºC for the solubilized enzyme and at 42.0ºC for the enzyme reconstituted in liposome. These transitions were also found by the FTIR technique. The experiments by DSC for the solubilized enzyme have shown the presence of three peaks at 54.7ºC, 64.7ºC and 67.8ºC. As for the reconstituted enzyme, transitions in lower temperatures between 30ºC and 40ºC (concerning the lipids) and also the preservation of the transition peak for the protein at 68.0ºC were observed. The Tryptophane fluorescence analysis for both enzyme forms has revealed emission maximum peak shifts starting from 60ºC. The Guaniddine presence has shown two transition points at 3 and 5 mol.L-1 for the solubilized Na,K-ATPase. The effect of different buffer media has shown that the enzyme presents higher contents in -helix at pH 7.5, concomitant with an increase of the intensity of tryptophane fluorescence emission in the pH range of 5.0 to 8.5. Analyzing all the techniques together we can propose a dissociation/denaturation mechanism in function of the temperature. First, the enzyme goes from its oligomeric ()2 state and forms protomers. The ATPase activity é totally lost ( over 60ºC) when the subunits are completely separated, when an subunits aggregation then occurs, through the cytoplasmatic domains. Finally, the analysis of the enzyme in different proportions of protein:detergent reveals that the NA,K-ATPase, in the presence of concentrations bellow CMS, is in the ()2 or yet in the ()4 form (Depending on protein concentration). Now for concentrations above CMS, the separation of the subunits occurs and consequent catalytic activity loss. Due to the ATPase activity dependence on its conformational form and oligomerization state, this study done with biophysical and biochemistry techniques, results in new information on the comprehension of the mechanisms that control the association processes, which is important to the enzyme function in the natural membrane.

A Na,K-ATPase é uma proteína encontrada na membrana plasmática de praticamente todas as células animais, que utiliza a energia derivada da hidrólise do ATP para transportar 3 íons Na+ e 2 íons K+. É composta por duas subunidades denominadas e . Um aspecto que ainda gera controvérsias se refere à sua forma de associação nativa e funcional como um protômero ou ainda na forma de oligômeros ()2 ou ()4. Uma forma de estudar essa enzima é pela sua solubilização da membrana, e posteriormente reconstituição em lipossomos de DPPC:DPPE. A caracterização cinética e estrutural desse sistema mostra que a enzima se apresenta na forma oligomérica ()2. O objetivo desse trabalho foi avaliar os mecanismos de dissociação e de desnaturação da Na,K-ATPase solubilizada bem como da reconstituída em lipossomos de DPPC:DPPE, por agentes físicos (temperatura) e químicos (relação proteína:detergente, uso de agentes caotrópicos como a guanidina e mudanças de pH), para interpretar as suas formas de associação e regulação. Para isso, foram realizados experimentos de dicroísmo circular (CD), calorimetria (DSC), infravermelho (FTIR), fluorescência de emissão do triptofano, tensão superficial, elasticidade, atividade catalítica (ATPase e pNPPase). Os estudos de CD em função da variação de temperatura mostraram que ocorre uma transição na curva de elipticidade (222 nm) a 43,7°C para a enzima solubilizada e a 42,0°C para a enzima reconstituída em lipossomos. Estas transições foram também encontradas pela técnica de FTIR. Os experimentos por DSC para a enzima solubilizada revelaram a presença de três picos em 54,7; 64,7 e 67,8°C. Já para a enzima reconstituída observam-se transições em menores temperaturas entre 30 a 40ºC (referentes aos lipídios) e ainda a preservação do pico de transição para proteína em 68,0°C. A análise de fluorescência de triptofano para ambas formas de enzima revelou deslocamentos de pico máximo de emissão a partir de 60°C. Já a presença de guanidina mostrou dois pontos de transição em 3 e 5 mol.L-1 para a Na,K-ATPase solubilizada. O efeito de diferentes meios tamponantes revelou que a enzima apresenta maiores conteúdos em -hélice em pH 7,5, concomitante com um aumento na intensidade de emissão de fluorescência do triptofano na faixa de pH de 5,0 a 8,5. Analisando conjuntamente todas as técnicas podemos propor um mecanismo de dissociação/desnaturação da enzima em função da temperatura. Primeiramente a enzima passa do seu estado oligomérico ()2 e forma protômeros . A atividade ATPase é perdida completamente (acima de 60ºC) quando as subunidades são completamente separadas, ocorrendo então uma agregação das subunidades , através dos domínios citoplasmáticos. Finalmente, a análise da enzima em diferentes proporções de proteína:detergente revela que a Na,K-ATPase, na presença de concentrações abaixo da CMC, se encontra na forma ()2 ou ainda ()4 (dependendo da concentração de proteína). Já para concentrações acima da CMC ocorre a separação das subunidades e consequente perda de atividade catalítica. Devido à dependência da atividade ATPase com seu estado conformacional e seu estado de oligomerização, este estudo realizado por técnicas bioquímicas e biofísicas, resulta em novas informações acerca da compreensão dos mecanismos que controlam o processo de associação, o qual é importante para a função da enzima na membrana natural. / Na,K-ATPase is a protein found in the plasmatic membrane of almost all animal cells and it uses the energy from ATP hydrolysis to transport 3 Na+ ions and 2 K+ ions. It is formed by subunits called and. One controversial aspect refers to its native and functional association form as a protomer or still in ()2 or ()4 oligomers form. One way to study this protein in our laboratory is by its solubilization from membrane, and later reconstitution in liposome from DPPC:DPPE. The kinetic and structural characterization fo this system shows that the enzyme presents itself in the oligomeric form ()2. The aim of this work was to evaluate the dissociation and denaturation mechanisms of the solubilized NA,K-ATPase as well as the one reconstituted in DPPC:DPPE liposome, by physic (temperature) and chemical agents (relation protein:detergent, use of chaotropic agents as Guanidine chloride, or still by the pH changes, to interpret its association and regulation forms. To that end, experiments of circular dichroism (CD), calorimetry (DSC), superficial tension, elasticity , catalytic activity (ATPase an pNPPase) were done. The CD studies in function of temperature variation have shown that a transition occurs in the ellipticity curve (222 nm) at 43.7ºC for the solubilized enzyme and at 42.0ºC for the enzyme reconstituted in liposome. These transitions were also found by the FTIR technique. The experiments by DSC for the solubilized enzyme have shown the presence of three peaks at 54.7ºC, 64.7ºC and 67.8ºC. As for the reconstituted enzyme, transitions in lower temperatures between 30ºC and 40ºC (concerning the lipids) and also the preservation of the transition peak for the protein at 68.0ºC were observed. The Tryptophane fluorescence analysis for both enzyme forms has revealed emission maximum peak shifts starting from 60ºC. The Guaniddine presence has shown two transition points at 3 and 5 mol.L-1 for the solubilized Na,K-ATPase. The effect of different buffer media has shown that the enzyme presents higher contents in -helix at pH 7.5, concomitant with an increase of the intensity of tryptophane fluorescence emission in the pH range of 5.0 to 8.5. Analyzing all the techniques together we can propose a dissociation/denaturation mechanism in function of the temperature. First, the enzyme goes from its oligomeric ()2 state and forms protomers. The ATPase activity é totally lost ( over 60ºC) when the subunits are completely separated, when an subunits aggregation then occurs, through the cytoplasmatic domains. Finally, the analysis of the enzyme in different proportions of protein:detergent reveals that the NA,K-ATPase, in the presence of concentrations bellow CMS, is in the ()2 or yet in the ()4 form (Depending on protein concentration). Now for concentrations above CMS, the separation of the subunits occurs and consequent catalytic activity loss. Due to the ATPase activity dependence on its conformational form and oligomerization state, this study done with biophysical and biochemistry techniques, results in new information on the comprehension of the mechanisms that control the association processes, which is important to the enzyme function in the natural membrane.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第17905号 / 農博第2028号 / 新制||農||1018(附属図書館) / 学位論文||H25||N4801(農学部図書室) / 30725 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 植田 和光, 教授 加納 健司, 教授 小川 順 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

The inhibitory effects on Escherichia coli ATPase activity by two venom peptides, lasioglossin II and mastoparan B. Membrane bound F1FO ATP synthase was isolated from E. coli strain pBWU13.4/DK8 and treated with varied concentrations of lasioglossin II and mastoparan B. Lasioglossin II caused very low inhibition of ATPase activity, but the inhibition profile of mastoparan B was suggestive of an interesting biological effect. A relatively shorter total length, a smaller net positive charge, and a reduced amphipathic character of both peptides, as compared to previously tested antimicrobial peptides, may account for the limited degree of inhibition observed in the present study.

ATP is the main cellular energy generated by the enzyme ATP synthase in almost all organisms from bacteria to vertebrates. While malfunction of the ATP synthase complex is responsible for several disease conditions, the enzyme itself can be used as a potent molecular drug target to combat many diseases including microbial infections, cancer, tuberculosis, and obesity. Recent widespread escalation of antibiotic resistant microbes in general and E. coli in particular demands novel alternative approaches to combat microbial infections. Inhibition of ATP synthase by inhibitors such as peptides is known to deprive microbes of required energy, resulting in microbial cell death. Therefore, we have examined the venom peptide induced inhibition of E. coli ATP synthase. It was found that venom peptides completely inhibited E. coli ATP synthase and the process of inhibition was found to be fully reversible. This study also links the antimicrobial properties of peptides in part to the inhibition of ATP synthase. Thus, selective use of ATP synthase as a molecular drug may have an important impact on biology and medicine.

Kinesins are molecular motors that transport intracellular cargos along microtubules (MTs) and influence the organization and dynamics of the MT cytoskeleton. Their force-generating functions arise from conformational changes in their motor domain as ATP is bound and hydrolyzed, and products are released. In the budding yeast Saccharomyces cerevisiae, the Kar3 kinesin forms heterodimers with one of two non-catalytic kinesin-like proteins, Cik1 and Vik1, which lack the ability to bind ATP, and yet they retain the capacity to bind MTs. Cik1 and Vik1 also influence and respond to the MT-binding and nucleotide states of Kar3, and differentially regulate the functions of Kar3 during yeast mating and mitosis. The mechanism by which Kar3/Cik1 and Kar3/Vik1 dimers operate remains unknown, but has important implications for understanding mechanical coordination between subunits of motor complexes that traverse cytoskeletal tracks. In this study, we show that the opportunistic human fungal pathogen Candida albicans (Ca) harbors a single version of this unique form of heterodimeric kinesin and we present the first in vitro characterization of this motor. Like its budding yeast counterpart, the Vik1-like subunit binds directly to MTs and strengthens the MT-binding affinity of the heterodimer. However, in contrast to ScKar3/Cik1 and ScKar3/Vik1, CaKar3/Vik1 exhibits weaker overall MT-binding affinity and lower ATPase activity. Preliminary investigations using a multiple motor motility assay indicate CaKar3/Vik1 may not be motile. Using a maltose binding protein tagging system, we determined the X-ray crystal structure of the CaKar3 motor domain and observed notable differences in its nucleotide-binding pocket relative to ScKar3 that appear to represent a previously unobserved state of the active site. Together, these studies broaden our knowledge of novel kinesin motor assemblies and shed new light on structurally dynamic regions of Kar3/Vik1-like motor complexes that help mediate mechanical coordination of its subunits. / Thesis (Master, Biochemistry) -- Queen's University, 2012-02-29 17:15:03.654

Dissertação(mestrado) - Universidade Federal do Rio Grande, Programa de Pós–Graduação em Aqüicultura, Instituto de Oceanografia, 2011. / Submitted by Cristiane Silva (cristiane_gomides@hotmail.com) on 2012-08-11T17:30:34Z No. of bitstreams: 1 dissertao mestrado_iuri salim abou anni.pdf: 920616 bytes, checksum: e1ed6eb173ad9593484417ec03fdce7b (MD5) / Approved for entry into archive by Bruna Vieira(bruninha_vieira@ibest.com.br) on 2012-12-19T14:26:35Z (GMT) No. of bitstreams: 1 dissertao mestrado_iuri salim abou anni.pdf: 920616 bytes, checksum: e1ed6eb173ad9593484417ec03fdce7b (MD5) / Made available in DSpace on 2012-12-19T14:26:35Z (GMT). No. of bitstreams: 1 dissertao mestrado_iuri salim abou anni.pdf: 920616 bytes, checksum: e1ed6eb173ad9593484417ec03fdce7b (MD5) Previous issue date: 2011 / O objetivo deste estudo foi avaliar os efeitos da salinidade sobre o comportamento iono e osmorregulatório, bem como o crescimento de juvenis de pampo Trachinotus marginatus. Um experimento foi realizado para estimar o ponto isosmótico e as concentrações iônicas plasmáticas do pampo. Os peixes (144,7 ± 28,4 g e 19,6 ± 2,0 cm) foram aclimatados durante quinze dias nas salinidades 4, 8, 12, 16 e 20 e amostras de sangue foram coletadas para análise da composição iônica e osmolalidade plasmática. O ponto isosmótico foi determinado pela regressão linear entre a osmolalidade plasmática e a osmolalidade da água. Em um segundo experimento, 320 peixes (2,14 ± 0,29 g e 5,11 ± 0,33 cm) foram aleatoriamente distribuídos em 16 tanques (50L). Cada tratamento foi mantido nas salinidades 3, 6, 12 e 32 (quatro repetições cada), equivalente a 25, 50, 100 e 267% do ponto de isosmótico. Durante o período experimental (28 dias), os peixes foram mantidos a 28 °C, pH 8,0, alcalinidade 135 mg CaCO3/L e saturação de oxigênio sempre superior a 90%. O consumo de oxigênio foi medido em cada salinidade. O segundo arco branquial de 12 peixes foi coletado para análise da atividade da enzima Na+/K+-ATPase. O ponto isosmótico do pampo foi determinado em 357,5 mOsmoles/kg H2O-1, o que equivale à salinidade 13,1. A osmolalidade plasmática, o hematócrito, a glicemia, o índice hepatossomático e atividade da Na+/K+-ATPase branquial não foram afetados pela salinidade. No segundo experimento, a maior taxa de consumo de oxigênio foi observado para os peixes criados na salinidade 3, enquanto a atividade da Na+/K+-ATPase branquial foi significativamente maior nesta salinidade em relação às salinidades 12 e 32. A concentração de glicogênio hepático da salinidade 3 foi significativamente menor em relação a salinidade 32. A atividade da tripsina no intestino e a umidade dos músculos não apresentaram variação significativa entre os tratamentos. O maior crescimento foi observado nas salinidades 3, 6 e 12. / The objective of this study was to evaluate the effect of salinity on juvenile growth of juvenile pompano Trachinotus marginatus. One experiment was done to estimate the isosmotic point of pompano, fish (144,7 ± 28,4 g e 19,6 ± 2,0 cm) were acclimated fortnightly at salinities 4, 8, 12, 16 and 20 and blood was sampled for osmolarity analysis. The isosmotic point was determined by linear regression between plasma osmolality and water osmolality. Later, 320 fish (2,1 ± 0,2 g and 5,1 ± 0,3 cm) were randomly distributed into 16 tanks (50L). Each treatment was maintained at salinities 3, 6, 12, and 32 (four replicates each), equivalent to 25, 50, 100 and 267% of the isosmotic point. During the experimental period (28 days), fish were maintained at 28°C, pH 8.0, alkalinity 135 mg/L of CaCO3, and oxygen saturation was always above 90%. Oxygen consumption was measured at each salinity. The second gill arch of 12 fish was collected for analysis of the Na+/K+-ATPase. The isosmotic point of T. marginatus was determined in 357,5 mOsmol/kg H2O-1, which is equivalent to salinity 13,1. Plasma osmolality, hematocrit, glycemia, hepatosomatic index and activity of Na+/K+-ATPase were not affected by salinity. In the second experiment, the highest rate of oxygen consumption was observed for fish reared at salinity 3, while the activity of Na+/K+-ATPase was significantly higher in salinity 3 in relation to salinity 12 and 32. The hepatic glycogen concentration of salinity 3 was significantly lower than the salinity 32. The activity of trypsin in the intestine and muscle humidity showed no significant variation between treatments. The best growth performance was observed at salinity 3, 6 and 12.