A rapid and inexpensive method to assay transport of short chain peptides across intestinal brush-border membrane vesicles from the European eel (Anguilla anguilla)
| Autor: | Carlo Storelli, Ivar Rønnestad, Amilcare Barca, Snorre Bakke, Antonio Danieli, Michele Maffia, Alessandro Romano, Tiziano Verri |
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| Přispěvatelé: | Verri, Tiziano, Danieli, A., Bakke, S., Romano, A., Barca, A., Ronnestad, I., Maffia, Michele, Storelli, Carlo |
| Rok vydání: | 2008 |
| Předmět: | |
| Zdroj: | Aquaculture Nutrition. 14:341-349 |
| ISSN: | 1365-2095 1353-5773 |
| DOI: | 10.1111/j.1365-2095.2007.00538.x |
| Popis: | Membrane potential depolarization due to electrogenic peptide transport activity was examined in eel (Anguilla anguilla) intestinal brush-border membrane vesicles (BBMV) by monitoring the fluorescence quenching of the voltage-sensitive dye 3,3'-diethylthiadicarbocyanine iodide. Our experimental approach consisted of generating an internal negative membrane potential mimicking in vivo conditions and measuring membrane potential depolarization due to different extravesicular dipeptides. Peptide-dependent membrane potential depolarization was observed in both the presence and absence of extravesicular Na+ and was inhibited by diethylpyrocarbonate, which is consistent with the involvement of electrogenic, Na+-independent, H+-dependent peptide transport activity. Kinetic analysis indicated that peptide-dependent membrane potential depolarization is a saturable process (K-m,K-app similar to 1.5 mmol L-1) and that within the 0.1-10 mmol L-1 peptide range a single carrier system is involved in the transport process. Our results suggest that a peptide transport activity, kinetically resembling the PepT1(Slc15A1)-type-mediated H+/peptide cotransport action, can be monitored in eel intestinal BBMV using an easy and inexpensive fluorescence assay. |
| Databáze: | OpenAIRE |
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