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Erythrocytes L-arginine y+ transporter inhibition by N-ethylmaleimide in ice-bath.
Cell Biochem Biophys. 2010 Nov; 58(2):69-73.CB

Abstract

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.

Authors+Show Affiliations

Programa de Pós-graduação em Medicina e Ciências da Saúde (Nefrologia), Faculdade de Medicina/Instituto de Pesquisas Biomédicas/Hospital São Lucas, Pontifícia Universidade Católica do Rio Grande do Sul, 2° Andar - Laboratório de Nefrologia, Instituto de Pesquisas Biomédicas/HSL, Porto Alegre, RS 90610-000, Brazil. bart@pucrs.brNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20668961

Citation

Pinheiro da Costa, Bartira Ercília, et al. "Erythrocytes L-arginine Y+ Transporter Inhibition By N-ethylmaleimide in Ice-bath." Cell Biochemistry and Biophysics, vol. 58, no. 2, 2010, pp. 69-73.
Pinheiro da Costa BE, de Almeida PB, Conceição IR, et al. Erythrocytes L-arginine y+ transporter inhibition by N-ethylmaleimide in ice-bath. Cell Biochem Biophys. 2010;58(2):69-73.
Pinheiro da Costa, B. E., de Almeida, P. B., Conceição, I. R., Antonello, I. C., d'Avila, D. O., & Poli-de-Figueiredo, C. E. (2010). Erythrocytes L-arginine y+ transporter inhibition by N-ethylmaleimide in ice-bath. Cell Biochemistry and Biophysics, 58(2), 69-73. https://doi.org/10.1007/s12013-010-9089-9
Pinheiro da Costa BE, et al. Erythrocytes L-arginine Y+ Transporter Inhibition By N-ethylmaleimide in Ice-bath. Cell Biochem Biophys. 2010;58(2):69-73. PubMed PMID: 20668961.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Erythrocytes L-arginine y+ transporter inhibition by N-ethylmaleimide in ice-bath. AU - Pinheiro da Costa,Bartira Ercília, AU - de Almeida,Priscilla Barcellos, AU - Conceição,Ioná Rosine, AU - Antonello,Ivan Carlos Ferreira, AU - d'Avila,Domingos O, AU - Poli-de-Figueiredo,Carlos Eduardo, PY - 2010/7/30/entrez PY - 2010/7/30/pubmed PY - 2011/1/29/medline SP - 69 EP - 73 JF - Cell biochemistry and biophysics JO - Cell Biochem Biophys VL - 58 IS - 2 N2 - 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. SN - 1559-0283 UR - https://www.unboundmedicine.com/medline/citation/20668961/Erythrocytes_L_arginine_y+_transporter_inhibition_by_N_ethylmaleimide_in_ice_bath_ L2 - https://dx.doi.org/10.1007/s12013-010-9089-9 DB - PRIME DP - Unbound Medicine ER -