Specificity of intestinal brush-border proline transport: cyanine dye studies.
The ability of rabbit jejunal brush borders to transport inhibitors of the imino carrier was investigated in membrane vesicles by measuring their ability to depolarize the membrane potential. Membrane potentials were monitored using a voltage-sensitive cyanine dye. Piperidine and pyrrolidine carboxylic acids, which are potent inhibitors of Na+-dependent proline transport (Ki less than 0.5 mM) depolarize the potential in a Na+-dependent, saturable manner indicating transport. On the other hand, N-methylated amino acids, which are fair inhibitors (Ki 2-10 mM), do not depolarize the membrane to any significant extent, but they competitively inhibit the L-proline transport signal. This indicates that these analogs are nontransported inhibitors of the imino carrier. The poor inhibitors niacin and pipolinic acid (Ki greater than 60 mM) depolarize the membrane about twice as much as proline and with low Kf values. This suggests separate carriers for these substrates.
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D052160:Benzothiazoles; D001692:Biological Transport; D002232:Carbocyanines; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism*; D007583:Jejunum / Q000378:metabolism; D007700:Kinetics; D008564:Membrane Potentials; D008871:Microvilli / Q000378:metabolism; D011392:Proline / Q000378:metabolism*; D011804:Quinolines; D011817:Rabbits
D000596:Amino Acids; D052160:Benzothiazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D011804:Quinolines; C012944:3,3'-dipropyl-2,2'-thiadicarbocyanine; D011392:Proline