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   <subfield code="a">Erythrocytes l -Arginine y+ Transporter Inhibition by N-Ethylmaleimide in Ice-bath</subfield>
   <subfield code="h">[Elektronische Daten]</subfield>
   <subfield code="c">[Bartira Pinheiro da Costa, Priscilla de Almeida, Ioná Conceição, Ivan Antonello, Domingos d'Avila, Carlos Poli-de-Figueiredo]</subfield>
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   <subfield code="a">Erythrocytes l-arginine uptake is conveyed by y+ and y+L membrane transport systems. Pre-incubation with N-ethylmaleimide for 10min at 37°C inhibits the y+ system. The aim of this study was to determine the ideal pre-incubation temperature in evaluating y+ and y+L systems. Cells were pre-incubated with or without N-ethylmaleimide for 10min at 4°C and 37°C. l-Arginine uptake was quantified by radioisotope and standard erythrocytes membrane flux methodology. Results demonstrate that erythrocytes l-arginine content is depleted by pre-incubation at 37°C for 10min, thus changing the V max measurement. The inhibitory effect of N-ethylmaleimide pre-incubation was temperature independent and already complete after 1min of incubation. No significant difference in kinetic parameters was detected between cells pre-incubated at 37°C or 4°C, under zero-trans conditions. In conclusion, we suggest that measurement of erythrocytes l-arginine uptake by y+ and y+L systems could be carried out without N-ethylmaleimide pre-incubation at 37°C.</subfield>
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