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Inhibition of plasminogen activator inhibitor-1 binding to endocytosis receptors of the low-density-lipoprotein receptor family by a peptide isolated from a phage display library

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Inhibition of plasminogen activator inhibitor-1 binding to endocytosis receptors of the low-density-lipoprotein receptor family by a peptide isolated from a phage display library. / Jensen, Jan Kristian; Malmendal, Anders; Schiøtt, Birgit; Skeldal, Sune; Pedersen, Katrine E; Celik, Leyla; Nielsen, Niels Chr.; Andreasen, Peter A; Wind, Troels.

I: Biochemical Journal, Bind 399, Nr. 3, 01.11.2006, s. 387-96.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Author

Jensen, Jan Kristian ; Malmendal, Anders ; Schiøtt, Birgit ; Skeldal, Sune ; Pedersen, Katrine E ; Celik, Leyla ; Nielsen, Niels Chr. ; Andreasen, Peter A ; Wind, Troels. / Inhibition of plasminogen activator inhibitor-1 binding to endocytosis receptors of the low-density-lipoprotein receptor family by a peptide isolated from a phage display library. I: Biochemical Journal. 2006 ; Bind 399, Nr. 3. s. 387-96.

Bibtex

@article{77224717156e4b468ef7c91f7e891b62,
title = "Inhibition of plasminogen activator inhibitor-1 binding to endocytosis receptors of the low-density-lipoprotein receptor family by a peptide isolated from a phage display library",
abstract = "The functions of the serpin PAI-1 (plasminogen activator inhibitor-1) are based on molecular interactions with its target proteases uPA and tPA (urokinase-type and tissue-type plasminogen activator respectively), with vitronectin and with endocytosis receptors of the low-density-lipoprotein family. Understanding the significance of these interactions would be facilitated by the ability to block them individually. Using phage display, we have identified the disulfide-constrained peptide motif CFGWC with affinity for natural human PAI-1. The three-dimensional structure of a peptide containing this motif (DVPCFGWCQDA) was determined by liquid-state NMR spectroscopy. A binding site in the so-called flexible joint region of PAI-1 was suggested by molecular modelling and validated through binding studies with various competitors and site-directed mutagenesis of PAI-1. The peptide with an N-terminal biotin inhibited the binding of the uPA-PAI-1 complex to the endocytosis receptors low-density-lipoprotein-receptor-related protein 1A (LRP-1A) and very-low-density-lipoprotein receptor (VLDLR) in vitro and inhibited endocytosis of the uPA-PAI-1 complex in U937 cells. We conclude that the isolated peptide represents a novel approach to pharmacological interference with the functions of PAI-1 based on inhibition of one specific molecular interaction.",
keywords = "Amino Acid Motifs, Amino Acid Sequence, Binding Sites, Binding, Competitive, Biotinylation, Cell Line, Cell Line, Tumor, Crystallography, X-Ray, Cystine, Endocytosis, Fibrosarcoma, Humans, Kidney, LDL-Receptor Related Protein 1, Micelles, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Neoplasm Proteins, Nuclear Magnetic Resonance, Biomolecular, Oligopeptides, Peptide Library, Plasminogen Activator Inhibitor 1, Protein Binding, Protein Conformation, Receptors, LDL, U937 Cells, Urokinase-Type Plasminogen Activator",
author = "Jensen, {Jan Kristian} and Anders Malmendal and Birgit Schi{\o}tt and Sune Skeldal and Pedersen, {Katrine E} and Leyla Celik and Nielsen, {Niels Chr.} and Andreasen, {Peter A} and Troels Wind",
year = "2006",
month = "11",
day = "1",
doi = "10.1042/BJ20060533",
language = "English",
volume = "399",
pages = "387--96",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "3",

}

RIS

TY - JOUR

T1 - Inhibition of plasminogen activator inhibitor-1 binding to endocytosis receptors of the low-density-lipoprotein receptor family by a peptide isolated from a phage display library

AU - Jensen, Jan Kristian

AU - Malmendal, Anders

AU - Schiøtt, Birgit

AU - Skeldal, Sune

AU - Pedersen, Katrine E

AU - Celik, Leyla

AU - Nielsen, Niels Chr.

AU - Andreasen, Peter A

AU - Wind, Troels

PY - 2006/11/1

Y1 - 2006/11/1

N2 - The functions of the serpin PAI-1 (plasminogen activator inhibitor-1) are based on molecular interactions with its target proteases uPA and tPA (urokinase-type and tissue-type plasminogen activator respectively), with vitronectin and with endocytosis receptors of the low-density-lipoprotein family. Understanding the significance of these interactions would be facilitated by the ability to block them individually. Using phage display, we have identified the disulfide-constrained peptide motif CFGWC with affinity for natural human PAI-1. The three-dimensional structure of a peptide containing this motif (DVPCFGWCQDA) was determined by liquid-state NMR spectroscopy. A binding site in the so-called flexible joint region of PAI-1 was suggested by molecular modelling and validated through binding studies with various competitors and site-directed mutagenesis of PAI-1. The peptide with an N-terminal biotin inhibited the binding of the uPA-PAI-1 complex to the endocytosis receptors low-density-lipoprotein-receptor-related protein 1A (LRP-1A) and very-low-density-lipoprotein receptor (VLDLR) in vitro and inhibited endocytosis of the uPA-PAI-1 complex in U937 cells. We conclude that the isolated peptide represents a novel approach to pharmacological interference with the functions of PAI-1 based on inhibition of one specific molecular interaction.

AB - The functions of the serpin PAI-1 (plasminogen activator inhibitor-1) are based on molecular interactions with its target proteases uPA and tPA (urokinase-type and tissue-type plasminogen activator respectively), with vitronectin and with endocytosis receptors of the low-density-lipoprotein family. Understanding the significance of these interactions would be facilitated by the ability to block them individually. Using phage display, we have identified the disulfide-constrained peptide motif CFGWC with affinity for natural human PAI-1. The three-dimensional structure of a peptide containing this motif (DVPCFGWCQDA) was determined by liquid-state NMR spectroscopy. A binding site in the so-called flexible joint region of PAI-1 was suggested by molecular modelling and validated through binding studies with various competitors and site-directed mutagenesis of PAI-1. The peptide with an N-terminal biotin inhibited the binding of the uPA-PAI-1 complex to the endocytosis receptors low-density-lipoprotein-receptor-related protein 1A (LRP-1A) and very-low-density-lipoprotein receptor (VLDLR) in vitro and inhibited endocytosis of the uPA-PAI-1 complex in U937 cells. We conclude that the isolated peptide represents a novel approach to pharmacological interference with the functions of PAI-1 based on inhibition of one specific molecular interaction.

KW - Amino Acid Motifs

KW - Amino Acid Sequence

KW - Binding Sites

KW - Binding, Competitive

KW - Biotinylation

KW - Cell Line

KW - Cell Line, Tumor

KW - Crystallography, X-Ray

KW - Cystine

KW - Endocytosis

KW - Fibrosarcoma

KW - Humans

KW - Kidney

KW - LDL-Receptor Related Protein 1

KW - Micelles

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Mutagenesis, Site-Directed

KW - Neoplasm Proteins

KW - Nuclear Magnetic Resonance, Biomolecular

KW - Oligopeptides

KW - Peptide Library

KW - Plasminogen Activator Inhibitor 1

KW - Protein Binding

KW - Protein Conformation

KW - Receptors, LDL

KW - U937 Cells

KW - Urokinase-Type Plasminogen Activator

U2 - 10.1042/BJ20060533

DO - 10.1042/BJ20060533

M3 - Journal article

C2 - 16813566

VL - 399

SP - 387

EP - 396

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 3

ER -