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Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells
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| Zeitschriftentitel: | European Journal of Biochemistry |
|---|---|
| Personen und Körperschaften: | , , , |
| In: | European Journal of Biochemistry, 271, 2004, 10, S. 2012-2017 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias |
|---|---|
| author |
Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias |
| spellingShingle |
Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias European Journal of Biochemistry Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells Biochemistry |
| author_sort |
neumann, jana |
| spelling |
Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias 0014-2956 1432-1033 Wiley Biochemistry http://dx.doi.org/10.1111/j.1432-1033.2004.04114.x <jats:p>The interaction of the antibacterial phosphonodipeptide alafosfalin with mammalian H<jats:sup>+</jats:sup>/peptide cotransporters was studied in Caco‐2 cells, expressing the low‐affinity intestinal type peptide transporter 1 (PEPT1), and SKPT cells, expressing the high‐affinity renal type peptide transporter 2 (PEPT2). Alafosfalin strongly inhibited the uptake of [<jats:sup>14</jats:sup>C]glycylsarcosine with <jats:italic>K</jats:italic><jats:sub>i</jats:sub> values of 0.19 ± 0.01 m<jats:sc>m</jats:sc> and 0.07 ± 0.01 m<jats:sc>m</jats:sc> for PEPT1 and PEPT2, respectively. Saturation kinetic studies revealed that in both cell types alafosfalin affected only the affinity constant (<jats:italic>K</jats:italic><jats:sub>t</jats:sub>) but not the maximal velocity (<jats:italic>V</jats:italic><jats:sub>max</jats:sub>) of glycylsarcosine (Gly‐Sar) uptake. The inhibition constants and the competitive nature of inhibition were confirmed in Dixon‐type experiments. Caco‐2 cells and SKPT cells were also cultured on permeable filters: apical uptake and transepithelial apical to basolateral flux of [<jats:sup>14</jats:sup>C]Gly‐Sar across Caco‐2 cell monolayers were reduced by alafosfalin (3 m<jats:sc>m</jats:sc>) by 73%. In SKPT cells, uptake of [<jats:sup>14</jats:sup>C]Gly‐Sar but not flux was inhibited by 61%. We found no evidence for an inhibition of the basolateral to apical uptake or flux of [<jats:sup>14</jats:sup>C]Gly‐Sar by alafosfalin. Alafosfalin (3 m<jats:sc>m</jats:sc>) did not affect the apical to basolateral [<jats:sup>14</jats:sup>C]mannitol flux. Determined in an Ussing‐type experiment with Caco‐2 cells cultured in Snapwells™, alafosfalin increased the short‐circuit current through Caco‐2 cell monolayers. We conclude that alafosfalin interacts with both H<jats:sup>+</jats:sup>/peptide symporters and that alafosfalin is actively transported across the intestinal epithelium in a H<jats:sup>+</jats:sup>‐symport, explaining its oral availability. The results also demonstrate that dipeptides where the C‐terminal carboxyl group is substituted by a phosphonic function represent high‐affinity substrates for mammalian H<jats:sup>+</jats:sup>/peptide cotransporters.</jats:p> Transport of the phosphonodipeptide alafosfalin by the H<sup>+</sup>/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells European Journal of Biochemistry |
| doi_str_mv |
10.1111/j.1432-1033.2004.04114.x |
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Online Free |
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Chemie und Pharmazie |
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ElectronicArticle |
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Wiley, 2004 |
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Wiley, 2004 |
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0014-2956 1432-1033 |
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0014-2956 1432-1033 |
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English |
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neumann2004transportofthephosphonodipeptidealafosfalinbythehpeptidecotransporterspept1andpept2inintestinalandrenalepithelialcells |
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2004 |
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Wiley |
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European Journal of Biochemistry |
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49 |
| title |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_unstemmed |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_full |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_fullStr |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_full_unstemmed |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_short |
Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_sort |
transport of the phosphonodipeptide alafosfalin by the h<sup>+</sup>/peptide cotransporters pept1 and pept2 in intestinal and renal epithelial cells |
| topic |
Biochemistry |
| url |
http://dx.doi.org/10.1111/j.1432-1033.2004.04114.x |
| publishDate |
2004 |
| physical |
2012-2017 |
| description |
<jats:p>The interaction of the antibacterial phosphonodipeptide alafosfalin with mammalian H<jats:sup>+</jats:sup>/peptide cotransporters was studied in Caco‐2 cells, expressing the low‐affinity intestinal type peptide transporter 1 (PEPT1), and SKPT cells, expressing the high‐affinity renal type peptide transporter 2 (PEPT2). Alafosfalin strongly inhibited the uptake of [<jats:sup>14</jats:sup>C]glycylsarcosine with <jats:italic>K</jats:italic><jats:sub>i</jats:sub> values of 0.19 ± 0.01 m<jats:sc>m</jats:sc> and 0.07 ± 0.01 m<jats:sc>m</jats:sc> for PEPT1 and PEPT2, respectively. Saturation kinetic studies revealed that in both cell types alafosfalin affected only the affinity constant (<jats:italic>K</jats:italic><jats:sub>t</jats:sub>) but not the maximal velocity (<jats:italic>V</jats:italic><jats:sub>max</jats:sub>) of glycylsarcosine (Gly‐Sar) uptake. The inhibition constants and the competitive nature of inhibition were confirmed in Dixon‐type experiments. Caco‐2 cells and SKPT cells were also cultured on permeable filters: apical uptake and transepithelial apical to basolateral flux of [<jats:sup>14</jats:sup>C]Gly‐Sar across Caco‐2 cell monolayers were reduced by alafosfalin (3 m<jats:sc>m</jats:sc>) by 73%. In SKPT cells, uptake of [<jats:sup>14</jats:sup>C]Gly‐Sar but not flux was inhibited by 61%. We found no evidence for an inhibition of the basolateral to apical uptake or flux of [<jats:sup>14</jats:sup>C]Gly‐Sar by alafosfalin. Alafosfalin (3 m<jats:sc>m</jats:sc>) did not affect the apical to basolateral [<jats:sup>14</jats:sup>C]mannitol flux. Determined in an Ussing‐type experiment with Caco‐2 cells cultured in Snapwells™, alafosfalin increased the short‐circuit current through Caco‐2 cell monolayers. We conclude that alafosfalin interacts with both H<jats:sup>+</jats:sup>/peptide symporters and that alafosfalin is actively transported across the intestinal epithelium in a H<jats:sup>+</jats:sup>‐symport, explaining its oral availability. The results also demonstrate that dipeptides where the C‐terminal carboxyl group is substituted by a phosphonic function represent high‐affinity substrates for mammalian H<jats:sup>+</jats:sup>/peptide cotransporters.</jats:p> |
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| author | Neumann, Jana, Bruch, Mandy, Gebauer, Sabine, Brandsch, Matthias |
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| description | <jats:p>The interaction of the antibacterial phosphonodipeptide alafosfalin with mammalian H<jats:sup>+</jats:sup>/peptide cotransporters was studied in Caco‐2 cells, expressing the low‐affinity intestinal type peptide transporter 1 (PEPT1), and SKPT cells, expressing the high‐affinity renal type peptide transporter 2 (PEPT2). Alafosfalin strongly inhibited the uptake of [<jats:sup>14</jats:sup>C]glycylsarcosine with <jats:italic>K</jats:italic><jats:sub>i</jats:sub> values of 0.19 ± 0.01 m<jats:sc>m</jats:sc> and 0.07 ± 0.01 m<jats:sc>m</jats:sc> for PEPT1 and PEPT2, respectively. Saturation kinetic studies revealed that in both cell types alafosfalin affected only the affinity constant (<jats:italic>K</jats:italic><jats:sub>t</jats:sub>) but not the maximal velocity (<jats:italic>V</jats:italic><jats:sub>max</jats:sub>) of glycylsarcosine (Gly‐Sar) uptake. The inhibition constants and the competitive nature of inhibition were confirmed in Dixon‐type experiments. Caco‐2 cells and SKPT cells were also cultured on permeable filters: apical uptake and transepithelial apical to basolateral flux of [<jats:sup>14</jats:sup>C]Gly‐Sar across Caco‐2 cell monolayers were reduced by alafosfalin (3 m<jats:sc>m</jats:sc>) by 73%. In SKPT cells, uptake of [<jats:sup>14</jats:sup>C]Gly‐Sar but not flux was inhibited by 61%. We found no evidence for an inhibition of the basolateral to apical uptake or flux of [<jats:sup>14</jats:sup>C]Gly‐Sar by alafosfalin. Alafosfalin (3 m<jats:sc>m</jats:sc>) did not affect the apical to basolateral [<jats:sup>14</jats:sup>C]mannitol flux. Determined in an Ussing‐type experiment with Caco‐2 cells cultured in Snapwells™, alafosfalin increased the short‐circuit current through Caco‐2 cell monolayers. We conclude that alafosfalin interacts with both H<jats:sup>+</jats:sup>/peptide symporters and that alafosfalin is actively transported across the intestinal epithelium in a H<jats:sup>+</jats:sup>‐symport, explaining its oral availability. The results also demonstrate that dipeptides where the C‐terminal carboxyl group is substituted by a phosphonic function represent high‐affinity substrates for mammalian H<jats:sup>+</jats:sup>/peptide cotransporters.</jats:p> |
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| imprint | Wiley, 2004 |
| imprint_str_mv | Wiley, 2004 |
| institution | DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275 |
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| match_str | neumann2004transportofthephosphonodipeptidealafosfalinbythehpeptidecotransporterspept1andpept2inintestinalandrenalepithelialcells |
| mega_collection | Wiley (CrossRef) |
| physical | 2012-2017 |
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| series | European Journal of Biochemistry |
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| spelling | Neumann, Jana Bruch, Mandy Gebauer, Sabine Brandsch, Matthias 0014-2956 1432-1033 Wiley Biochemistry http://dx.doi.org/10.1111/j.1432-1033.2004.04114.x <jats:p>The interaction of the antibacterial phosphonodipeptide alafosfalin with mammalian H<jats:sup>+</jats:sup>/peptide cotransporters was studied in Caco‐2 cells, expressing the low‐affinity intestinal type peptide transporter 1 (PEPT1), and SKPT cells, expressing the high‐affinity renal type peptide transporter 2 (PEPT2). Alafosfalin strongly inhibited the uptake of [<jats:sup>14</jats:sup>C]glycylsarcosine with <jats:italic>K</jats:italic><jats:sub>i</jats:sub> values of 0.19 ± 0.01 m<jats:sc>m</jats:sc> and 0.07 ± 0.01 m<jats:sc>m</jats:sc> for PEPT1 and PEPT2, respectively. Saturation kinetic studies revealed that in both cell types alafosfalin affected only the affinity constant (<jats:italic>K</jats:italic><jats:sub>t</jats:sub>) but not the maximal velocity (<jats:italic>V</jats:italic><jats:sub>max</jats:sub>) of glycylsarcosine (Gly‐Sar) uptake. The inhibition constants and the competitive nature of inhibition were confirmed in Dixon‐type experiments. Caco‐2 cells and SKPT cells were also cultured on permeable filters: apical uptake and transepithelial apical to basolateral flux of [<jats:sup>14</jats:sup>C]Gly‐Sar across Caco‐2 cell monolayers were reduced by alafosfalin (3 m<jats:sc>m</jats:sc>) by 73%. In SKPT cells, uptake of [<jats:sup>14</jats:sup>C]Gly‐Sar but not flux was inhibited by 61%. We found no evidence for an inhibition of the basolateral to apical uptake or flux of [<jats:sup>14</jats:sup>C]Gly‐Sar by alafosfalin. Alafosfalin (3 m<jats:sc>m</jats:sc>) did not affect the apical to basolateral [<jats:sup>14</jats:sup>C]mannitol flux. Determined in an Ussing‐type experiment with Caco‐2 cells cultured in Snapwells™, alafosfalin increased the short‐circuit current through Caco‐2 cell monolayers. We conclude that alafosfalin interacts with both H<jats:sup>+</jats:sup>/peptide symporters and that alafosfalin is actively transported across the intestinal epithelium in a H<jats:sup>+</jats:sup>‐symport, explaining its oral availability. The results also demonstrate that dipeptides where the C‐terminal carboxyl group is substituted by a phosphonic function represent high‐affinity substrates for mammalian H<jats:sup>+</jats:sup>/peptide cotransporters.</jats:p> Transport of the phosphonodipeptide alafosfalin by the H<sup>+</sup>/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells European Journal of Biochemistry |
| spellingShingle | Neumann, Jana, Bruch, Mandy, Gebauer, Sabine, Brandsch, Matthias, European Journal of Biochemistry, Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells, Biochemistry |
| title | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_full | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_fullStr | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_full_unstemmed | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_short | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| title_sort | transport of the phosphonodipeptide alafosfalin by the h<sup>+</sup>/peptide cotransporters pept1 and pept2 in intestinal and renal epithelial cells |
| title_unstemmed | Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells |
| topic | Biochemistry |
| url | http://dx.doi.org/10.1111/j.1432-1033.2004.04114.x |