A rapid method for the functional reconstitution of amino acid transport systems from rat liver plasma membranes. Partial purification of System A.
A rapid method for the functional reconstruction of amino acid transport from liver plasma-membrane vesicles using the neutral detergent decanoyl-N-glucamide ('MEGA-10') is described. The method is a modification of that previously employed in this laboratory for reconstitution of amino acid transport systems from kidney brush-border membranes [Lynch & McGivan (1987) Biochem. J. 244, 503-508]. The transport activities termed 'System A', 'System N', and 'System L' are all reconstituted. The reconstitution procedure is rapid and efficient and is suitable as an assay for transport activity in studies involving membrane fractionation. By using this reconstitution procedure, System A transport activity was partially purified by lectin-affinity chromatography.
Quesada
A R
AR
Department of Biochemistry, University of Bristol, U.K.
McGivan
J D
JD
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D000621:Aminoisobutyric Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D007930:Leucine / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008627:Mercuric Chloride / Q000494:pharmacology; D008634:Mersalyl / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D000596:Amino Acids; D000621:Aminoisobutyric Acids; D008563:Membrane Lipids; D008565:Membrane Proteins; D013439:Sulfhydryl Reagents; D005973:Glutamine; C017911:2-(methylamino)isobutyric acid; D008627:Mercuric Chloride; D008634:Mersalyl; D007930:Leucine; D000409:Alanine