Reconstitution and identification of the major Na(+)-dependent neutral amino acid-transport protein from bovine renal brush-border membrane vesicles.
Amino acid transport activity from bovine renal brush-border membrane vesicles (BBMV) was reconstituted into phospholipid vesicles composed of phosphatidylcholine/5% stearylamine. Reconstitutable transport activity was enhanced in protein fractions binding to various lectins. When solubilized BBMV were fractionated on peanut lectin, a single protein band of average molecular mass 132 kDa was obtained. When this protein fraction was reconstituted into phospholipid membrane vesicles, amino acid transport activity was obtained with properties similar to those in native BBMV with regard to amino acid specificity, although the cation specificity was different. A monoclonal antibody which reacted with the same protein removed reconstitutable amino acid transport activity from solubilized BBMV. These findings may provide the first identification of a renal amino acid-transporting protein, although confirmation of this identification by other approaches will be required.
Doyle
F A
FA
Department of Biochemistry, School of Medical Sciences, University Walk, Bristol, U.K.
McGivan
J D
JD
D000409:Alanine / Q000378:metabolism*; D000588:Amines / Q000494:pharmacology; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008871:Microvilli / Q000378:metabolism*; D008970:Molecular Weight; D010713:Phosphatidylcholines; D051381:Rats; D012964:Sodium / Q000494:pharmacology*; D013379:Substrate Specificity
D000588:Amines; D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D008081:Liposomes; D010713:Phosphatidylcholines; D012964:Sodium; C009317:stearylamine; D000409:Alanine