Erythrocytes l-Arginine y+ Transporter Inhibition by N-Ethylmaleimide in Ice-bath

Erythrocytes l-Arginine y+ Transporter Inhibition by N-Ethylmaleimide in Ice-bath

Erythrocytes l-arginine uptake is conveyed by y+ and y+L membrane transport systems. Pre-incubation with N-ethylmaleimide for 10 min 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...

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Bibliographic Details
Published in:Cell biochemistry and biophysics Vol. 58; no. 2; pp. 69 - 73
Main Authors: Pinheiro da Costa, Bartira Ercília, de Almeida, Priscilla Barcellos, Conceição, Ioná Rosine, Antonello, Ivan Carlos Ferreira, d'Avila, Domingos O, Poli-de-Figueiredo, Carlos Eduardo
Format: Journal Article
Language:English
Published: New York New York : Humana Press Inc 01-11-2010
Humana Press Inc
Springer Nature B.V
Subjects:
Amino Acid Transport System y+ - antagonists & inhibitors
Amino Acid Transport System y+ - metabolism
Amino Acid Transport System y+L - antagonists & inhibitors
Amino Acid Transport System y+L - metabolism
Arginine - metabolism
Biochemistry
Biological and Medical Physics
Biological Transport - drug effects
Biomedical and Life Sciences
Biophysics
Biotechnology
Carrier
Cationic amino acid transporters
Cell Biology
Cytological Techniques - methods
Erythrocytes
Erythrocytes - drug effects
Erythrocytes - metabolism
Ethylmaleimide - pharmacology
Ice
Kinetics
Life Sciences
Membrane transporters
nitric oxide
Original Paper
Pharmacology/Toxicology
Temperature
Cationic amino acid transporters
Carrier
Membrane transporters
Nitric oxide
Erythrocytes
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Summary:Erythrocytes l-arginine uptake is conveyed by y+ and y+L membrane transport systems. Pre-incubation with N-ethylmaleimide for 10 min 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 10 min 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 10 min, thus changing the V max measurement. The inhibitory effect of N-ethylmaleimide pre-incubation was temperature independent and already complete after 1 min 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.
Bibliography:http://dx.doi.org/10.1007/s12013-010-9089-9
ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:1085-9195
1559-0283
DOI:10.1007/s12013-010-9089-9