Sodium-dependent amino acid transport is preserved in lyophilized reconstituted apical membranes from intestinal epithelium
| Autor: | Bruce R. Stevens, Robert L. Preston |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Male
Brush border Biophysics Biochemistry Valinomycin chemistry.chemical_compound medicine Animals Amino acid transporter Amino Acids Intestinal Mucosa Molecular Biology chemistry.chemical_classification Chromatography Microvilli Vesicle Sodium Biological Transport Cell Biology Amino acid Microscopy Electron Membrane Freeze Drying chemistry Active transport Mannitol Rabbits medicine.drug |
| Zdroj: | Analytical biochemistry. 265(1) |
| ISSN: | 0003-2697 |
| Popis: | We demonstrate for the first time that functional electrogenic Na(+)-dependent amino acid transport is preserved for extended periods when purified brush border membranes prepared in hypotonic media are lyophilized and then rehydrated in buffer containing mannitol, NaSCN, and/or KSCN/valinomycin. Reconstituted lyophilized apical membranes from small intestine formed morphologically, physiologically, and thermodynamically normal vesicles which transported L-alanine via system B into an osmotically active space energized by secondary active transport, as measured under equilibrium and nonequilibrium conditions. The lyophilized membranes are readily prepared and stored, thereby providing a means to pool large quantities of formed vesicles that are useful in examining cloned and reconstituted native amino acid transporter polypeptides. |
| Databáze: | OpenAIRE |
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