PMID: 45779Jan 1, 1978

Reconstitution of neutral amino acid transport system from Ehrlich ascites tumor cells

Membrane Biochemistry
G CecchiniD L Oxender

Abstract

Amino acid transport systems for alanine and leucine have been reconstituted into artificial lipid vesicles. Purified plasma membrane vesicles from Ehrlich ascites cells were dissolved in 2% sodium cholate, 1 mM dithiothreitol, 0.5 mM EDTA, a mixture which solubilized approximately 50% of the membrane protein. This solubilized protein fraction was further purified by a combination of ammonium sulfate precipitations, gel filtration, and DEAE-cellulose chromatography. A fraction containing approximately 15 Coomassie blue staining bands on sodium dodecyl sulfate gels was obtained. This material was reconstituted into liposomes, and preliminary results demonstrated transport of alanine and leucine dependent on a sodium gradient. In addition, an electrogenic gradient mediated by valinomycin-induced potassium diffusion seemed to stimulate alanine uptake further.

References

Jan 1, 1983·The Journal of Membrane Biology·R J Turner
May 27, 1980·Biochimica Et Biophysica Acta·I C West

Citations

Jun 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·R T Hamilton, M Nilsen-Hamilton
Dec 1, 1976·Journal of Cellular Physiology·R M Johnstone, C Bardin
Feb 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·M Kasahara, P C Hinkle
Jan 1, 1974·The Journal of Membrane Biology·M Colombini, R M Johnstone

Related Concepts

SDS-PAGE
Valinomycin
Uptake
Chromatography, DEAE-Cellulose
Dithiothreitol
Amino Acid Transporter
Abufne
Vesicle
Sodium Cholate
Coomassie blue

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