Phage display screening of epithelial cell monolayers treated with EGTA: identification of peptide FDFWITP that modulates tight junction activity.
Identifieur interne : 002B41 ( Main/Exploration ); précédent : 002B40; suivant : 002B42Phage display screening of epithelial cell monolayers treated with EGTA: identification of peptide FDFWITP that modulates tight junction activity.
Auteurs : Richard E. Herman [États-Unis] ; Ekaterina G. Makienko ; Mary G. Prieve ; Mark Fuller ; Michael E. Houston ; Paul H. JohnsonSource :
- Journal of biomolecular screening [ 1087-0571 ] ; 2007.
Descripteurs français
- KwdFr :
- Acide egtazique (pharmacologie), Animaux, Banque de peptides, Calcium (métabolisme), Cellules Caco-2, Cellules épithéliales (), Cellules épithéliales (cytologie), Cellules épithéliales (métabolisme), Cellules épithéliales (virologie), Chiens, Chélateurs (pharmacologie), Données de séquences moléculaires, Fluorescéine-5-isothiocyanate (métabolisme), Humains, Impédance électrique, Jonctions serrées (), Jonctions serrées (physiologie), Peptides (), Peptides (pharmacologie), Perméabilité (), Séquence d'acides aminés.
- MESH :
- cytologie : Cellules épithéliales.
- métabolisme : Calcium, Cellules épithéliales, Fluorescéine-5-isothiocyanate.
- pharmacologie : Acide egtazique, Chélateurs, Peptides.
- physiologie : Jonctions serrées.
- virologie : Cellules épithéliales.
- Animaux, Banque de peptides, Cellules Caco-2, Cellules épithéliales, Chiens, Données de séquences moléculaires, Humains, Impédance électrique, Jonctions serrées, Peptides, Perméabilité, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Caco-2 Cells, Calcium (metabolism), Chelating Agents (pharmacology), Dogs, Egtazic Acid (pharmacology), Electric Impedance, Epithelial Cells (cytology), Epithelial Cells (drug effects), Epithelial Cells (metabolism), Epithelial Cells (virology), Fluorescein-5-isothiocyanate (metabolism), Humans, Molecular Sequence Data, Peptide Library, Peptides (chemistry), Peptides (pharmacology), Permeability (drug effects), Tight Junctions (drug effects), Tight Junctions (physiology).
- MESH :
- chemical , chemistry : Peptides.
- chemical , metabolism : Calcium, Fluorescein-5-isothiocyanate.
- chemical , pharmacology : Chelating Agents, Egtazic Acid, Peptides.
- cytology : Epithelial Cells.
- drug effects : Epithelial Cells, Permeability, Tight Junctions.
- metabolism : Epithelial Cells.
- physiology : Tight Junctions.
- virology : Epithelial Cells.
- Amino Acid Sequence, Animals, Caco-2 Cells, Dogs, Electric Impedance, Humans, Molecular Sequence Data, Peptide Library.
Abstract
Phage display was used to screen for peptides that modulate the activity of epithelial cell tight junctions. Panning with a phage library that displays random 7-mers was performed using monolayers of human bronchial epithelial cells (16HBE14o(-)) treated with a calcium chelator, ethylene glycol-bis(2-aminoethylether)- N, N, N', N'-tetraacetic acid (EGTA), to increase accessibility to the junctional complex/paracellular space, followed by subtractive panning. A novel peptide, FDFWITP, identified as a potential tight junction modulator, was synthesized in linear and cyclic forms with lysine residues added to improve solubility. The cyclic form of the peptide reduced transepithelial electrical resistance (TER) in a concentration-dependent manner (80% reduction at 100 microM and 95% reduction at 500 microM) and was reversible within 2 h; the linear form only affected TER at the highest concentration. Interestingly, the constrained peptide did not increase permeation of the model small molecule, fluorescein. The highly selective activity of FDFWITP supports the hypothesis that ions and small molecules may be transported paracellularly across tight junctions by separate pathways.
DOI: 10.1177/1087057107310216
PubMed: 18040053
Affiliations:
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Le document en format XML
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Prieve</name></author><author><name sortKey="Fuller, Mark" sort="Fuller, Mark" uniqKey="Fuller M" first="Mark" last="Fuller">Mark Fuller</name></author><author><name sortKey="Houston, Michael E" sort="Houston, Michael E" uniqKey="Houston M" first="Michael E" last="Houston">Michael E. Houston</name></author><author><name sortKey="Johnson, Paul H" sort="Johnson, Paul H" uniqKey="Johnson P" first="Paul H" last="Johnson">Paul H. 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Herman</name><affiliation wicri:level="1"><nlm:affiliation>Nastech Pharmaceutical Company, Inc., Bothell, Washington 98021, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Nastech Pharmaceutical Company, Inc., Bothell, Washington 98021</wicri:regionArea><wicri:noRegion>Washington 98021</wicri:noRegion></affiliation></author><author><name sortKey="Makienko, Ekaterina G" sort="Makienko, Ekaterina G" uniqKey="Makienko E" first="Ekaterina G" last="Makienko">Ekaterina G. Makienko</name></author><author><name sortKey="Prieve, Mary G" sort="Prieve, Mary G" uniqKey="Prieve M" first="Mary G" last="Prieve">Mary G. Prieve</name></author><author><name sortKey="Fuller, Mark" sort="Fuller, Mark" uniqKey="Fuller M" first="Mark" last="Fuller">Mark Fuller</name></author><author><name sortKey="Houston, Michael E" sort="Houston, Michael E" uniqKey="Houston M" first="Michael E" last="Houston">Michael E. Houston</name></author><author><name sortKey="Johnson, Paul H" sort="Johnson, Paul H" uniqKey="Johnson P" first="Paul H" last="Johnson">Paul H. Johnson</name></author></analytic><series><title level="j">Journal of biomolecular screening</title><idno type="ISSN">1087-0571</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Caco-2 Cells</term><term>Calcium (metabolism)</term><term>Chelating Agents (pharmacology)</term><term>Dogs</term><term>Egtazic Acid (pharmacology)</term><term>Electric Impedance</term><term>Epithelial Cells (cytology)</term><term>Epithelial Cells (drug effects)</term><term>Epithelial Cells (metabolism)</term><term>Epithelial Cells (virology)</term><term>Fluorescein-5-isothiocyanate (metabolism)</term><term>Humans</term><term>Molecular Sequence Data</term><term>Peptide Library</term><term>Peptides (chemistry)</term><term>Peptides (pharmacology)</term><term>Permeability (drug effects)</term><term>Tight Junctions (drug effects)</term><term>Tight Junctions (physiology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acide egtazique (pharmacologie)</term><term>Animaux</term><term>Banque de peptides</term><term>Calcium (métabolisme)</term><term>Cellules Caco-2</term><term>Cellules épithéliales ()</term><term>Cellules épithéliales (cytologie)</term><term>Cellules épithéliales (métabolisme)</term><term>Cellules épithéliales (virologie)</term><term>Chiens</term><term>Chélateurs (pharmacologie)</term><term>Données de séquences moléculaires</term><term>Fluorescéine-5-isothiocyanate (métabolisme)</term><term>Humains</term><term>Impédance électrique</term><term>Jonctions serrées ()</term><term>Jonctions serrées (physiologie)</term><term>Peptides ()</term><term>Peptides (pharmacologie)</term><term>Perméabilité ()</term><term>Séquence d'acides aminés</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Peptides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Calcium</term><term>Fluorescein-5-isothiocyanate</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Chelating Agents</term><term>Egtazic Acid</term><term>Peptides</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Cellules épithéliales</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Epithelial Cells</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Epithelial Cells</term><term>Permeability</term><term>Tight Junctions</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Epithelial Cells</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Calcium</term><term>Cellules épithéliales</term><term>Fluorescéine-5-isothiocyanate</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Acide egtazique</term><term>Chélateurs</term><term>Peptides</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Jonctions serrées</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Tight Junctions</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules épithéliales</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Epithelial Cells</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Caco-2 Cells</term><term>Dogs</term><term>Electric Impedance</term><term>Humans</term><term>Molecular Sequence Data</term><term>Peptide Library</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Banque de peptides</term><term>Cellules Caco-2</term><term>Cellules épithéliales</term><term>Chiens</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Impédance électrique</term><term>Jonctions serrées</term><term>Peptides</term><term>Perméabilité</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Phage display was used to screen for peptides that modulate the activity of epithelial cell tight junctions. Panning with a phage library that displays random 7-mers was performed using monolayers of human bronchial epithelial cells (16HBE14o(-)) treated with a calcium chelator, ethylene glycol-bis(2-aminoethylether)- N, N, N', N'-tetraacetic acid (EGTA), to increase accessibility to the junctional complex/paracellular space, followed by subtractive panning. A novel peptide, FDFWITP, identified as a potential tight junction modulator, was synthesized in linear and cyclic forms with lysine residues added to improve solubility. The cyclic form of the peptide reduced transepithelial electrical resistance (TER) in a concentration-dependent manner (80% reduction at 100 microM and 95% reduction at 500 microM) and was reversible within 2 h; the linear form only affected TER at the highest concentration. Interestingly, the constrained peptide did not increase permeation of the model small molecule, fluorescein. The highly selective activity of FDFWITP supports the hypothesis that ions and small molecules may be transported paracellularly across tight junctions by separate pathways.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Fuller, Mark" sort="Fuller, Mark" uniqKey="Fuller M" first="Mark" last="Fuller">Mark Fuller</name><name sortKey="Houston, Michael E" sort="Houston, Michael E" uniqKey="Houston M" first="Michael E" last="Houston">Michael E. Houston</name><name sortKey="Johnson, Paul H" sort="Johnson, Paul H" uniqKey="Johnson P" first="Paul H" last="Johnson">Paul H. Johnson</name><name sortKey="Makienko, Ekaterina G" sort="Makienko, Ekaterina G" uniqKey="Makienko E" first="Ekaterina G" last="Makienko">Ekaterina G. Makienko</name><name sortKey="Prieve, Mary G" sort="Prieve, Mary G" uniqKey="Prieve M" first="Mary G" last="Prieve">Mary G. Prieve</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Herman, Richard E" sort="Herman, Richard E" uniqKey="Herman R" first="Richard E" last="Herman">Richard E. Herman</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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