Characterisation and Partial Purification of the Na+-Dependent Neutral Amino Acid Transporting System from Bovine Kidney Brush Border Membranes

2015 ◽  
pp. 37-42 ◽  
Author(s):  
J. D. McGivan ◽  
A. Lynch
Keyword(s):  
Amino Acid ◽  
Brush Border ◽  
Neutral Amino Acid ◽  
Partial Purification ◽  
Bovine Kidney ◽  
Brush Border Membranes

scholarly journals A rapid method for the reconstitution of Na+-dependent neutral amino acid transport from bovine renal brush-border membranes

1987 ◽  
Vol 244 (3) ◽  
pp. 503-508 ◽  
Author(s):  
A M Lynch ◽  
J D McGivan
Keyword(s):  
Amino Acid ◽  
Rapid Method ◽  
Brush Border ◽  
Amino Acid Transport ◽  
Neutral Amino Acid ◽  
Acid Transport ◽  
Transport Activity ◽  
Brush Border Membranes ◽  
Renal Brush Border Membranes ◽  
Renal Brush Border

1. A simple and rapid method for the reconstitution of Na+-dependent neutral amino acid transport activity from bovine renal brush border membranes is described. 2. The neutral detergent decanoyl-N-methylglucamide (‘MEGA-10’) was employed to solubilize the membrane protein. This obviated the necessity for a prolonged dialysis step. 3. The properties of amino acid transport in these vesicles were similar to those observed in native membranes. 4. This should be a useful procedure in the eventual identification and isolation of amino acid transport proteins.

Na+-dependent neutral amino acid transporter ATB0 is a rabbit epithelial cell brush-border protein

2001 ◽  
Vol 281 (3) ◽  
pp. C963-C971 ◽  
Author(s):  
Nelly E. Avissar ◽  
Charlotte K. Ryan ◽  
Vadivel Ganapathy ◽  
Harry C. Sax
Keyword(s):  
Amino Acid ◽  
Hela Cells ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Polyclonal Antibodies ◽  
Neutral Amino Acid ◽  
Rabbit Kidney ◽  
Western Blots ◽  
Amino Acid Transport System ◽  
Rabbit Tissues

System B0 activity accounts for the majority of intestinal and kidney luminal neutral amino acid absorption. An amino acid transport system, called ATB0 (also known as ASCT2), with functional characteristics similar to those of system B0, has been recently cloned. We generated polyclonal antibodies to human and rabbit ATB0 COOH-terminal peptides and used Western blot analysis to detect ATB0 protein in rabbit tissues, rabbit ileal brush-border membrane vesicles (BBMV), and HeLa cells transfected with plasmids containing ATB0 cDNAs. Immunohistochemistry was used to localize ATB0 in rabbit kidney and intestine. In Western blots of rabbit tissues, ATB0 was a broad smear of 78- to 85-kDa proteins. In transfected HeLa cells, ATB0 appeared as a smear consisting of 57- to 65-kDa proteins. The highest expression was found in the kidney. ATB0 was enriched in rabbit ileal BBMV and in HeLa cells transfected with ATB0 cDNAs. In the kidney and in the intestine, ATB0 was confined to the brush-border membrane (BBM) of the proximal tubular cell and of the enterocyte, respectively. Tissue and intracellular distribution of ATB0 protein parallels that of system B0 activity. ATB0protein could be the transporter responsible for system B0in the BBM of epithelial cells.

scholarly journals Different handling of parathyrin by basal-lateral and brush-border membranes of the bovine kidney cortex

1980 ◽  
Vol 188 (3) ◽  
pp. 649-656 ◽  
Author(s):  
H Mohr ◽  
R D Hesch
Keyword(s):  
Plasma Membrane ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Kidney Cortex ◽  
Small Peptides ◽  
Uniform Size ◽  
Bovine Kidney ◽  
Brush Border Membranes ◽  
Membrane Bound ◽  
Uniform Size Distribution

The two parts of the bovine kidney cortex plasma membrane, the basal-lateral and the brush-border membrane, were simultaneously prepared from the same organ. Both types of membrane bound parathyrin, but only from the basal-lateral fraction was the hormone displaceable by its bioactive N-terminal fragment. In parallel, parathyrin-stimulated adenylate cyclase was predominantly found in basal-lateral membranes. The hormone was fragmented by both membrane types. Basal-lateral membranes generated fragments with a rather uniform size distribution (somewhat smaller than the intact peptide) and apparently preferred the hormone itself as a substrate. In contrast, the fragments produced by brush-border membranes were numberous small peptides.

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