H+/peptide cotransport in brush-border membrane vesicles (BBMVs) from eel (Anguilla anguilla) intestine was studied by measuring D-[H-3]-phenylalanyl-L-alanine uptake and by monitoring peptide-dependent intravesicular acidification using the pH-sensitive dye Acridine Orange, D-[H-3]-phenylalanyl-L-alanine influx was greatly stimulated by an inside-negative membrane potential and enhanced by an inwardly directed H+ gradient. In parallel, vesicular Hf influx was significantly increased in the presence of extravesicular D-phenylalanyl-L-alanine or a series of glycyl and L-prolyl peptides. H+/peptide cotransport displayed saturable kinetics involving a single carrier system with apparent substrate affinities of 0.9-2.6 mmol l(-1) depending on the particular peptide. All substrates tested competed with this system. Pre-incubation of BBMVs,vith dipeptides prevented diethylpyrocarbonate inhibition of transport activity, suggesting that the substrates mask histidine residues involved in the catalytic function of the transporter. Using human PepT1-specific primers, a reverse transcriptisn-polymerase chain reaction (RT-PCR) signal was detected in eel intestine. Our results suggest that, in eel intestine, a brush-border membrane 'low-affinity'-type H+/peptide cotransport system is present that shares kinetic features with the mammalian intestinal PepT1-type transporters.

Characterization of the H+/peptide cotransporter of eel intestinal brush-border membranes.

VERRI, Tiziano;MAFFIA, Michele;DANIELI, Antonio;STORELLI, Carlo
2000-01-01

Abstract

H+/peptide cotransport in brush-border membrane vesicles (BBMVs) from eel (Anguilla anguilla) intestine was studied by measuring D-[H-3]-phenylalanyl-L-alanine uptake and by monitoring peptide-dependent intravesicular acidification using the pH-sensitive dye Acridine Orange, D-[H-3]-phenylalanyl-L-alanine influx was greatly stimulated by an inside-negative membrane potential and enhanced by an inwardly directed H+ gradient. In parallel, vesicular Hf influx was significantly increased in the presence of extravesicular D-phenylalanyl-L-alanine or a series of glycyl and L-prolyl peptides. H+/peptide cotransport displayed saturable kinetics involving a single carrier system with apparent substrate affinities of 0.9-2.6 mmol l(-1) depending on the particular peptide. All substrates tested competed with this system. Pre-incubation of BBMVs,vith dipeptides prevented diethylpyrocarbonate inhibition of transport activity, suggesting that the substrates mask histidine residues involved in the catalytic function of the transporter. Using human PepT1-specific primers, a reverse transcriptisn-polymerase chain reaction (RT-PCR) signal was detected in eel intestine. Our results suggest that, in eel intestine, a brush-border membrane 'low-affinity'-type H+/peptide cotransport system is present that shares kinetic features with the mammalian intestinal PepT1-type transporters.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11587/300120
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