RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1

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RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1. / Madsen, Jeppe B; Dupont, Daniel Miotto; Andersen, Thomas B; Nielsen, Anne F; Sang, Lu; Brix, Ditte M; Jensen, Jan K; Broos, Thomas; Hendrickx, Maarten L V; Christensen, Anni; Kjems, Jørgen; Andreasen, Peter A.

I: Biochemistry, Bind 49, Nr. 19, 18.05.2010, s. 4103-15.

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

Harvard

Madsen, JB, Dupont, DM, Andersen, TB, Nielsen, AF, Sang, L, Brix, DM, Jensen, JK, Broos, T, Hendrickx, MLV, Christensen, A, Kjems, J & Andreasen, PA 2010, 'RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1', Biochemistry, bind 49, nr. 19, s. 4103-15. https://doi.org/10.1021/bi100066j

APA

Madsen, J. B., Dupont, D. M., Andersen, T. B., Nielsen, A. F., Sang, L., Brix, D. M., Jensen, J. K., Broos, T., Hendrickx, M. L. V., Christensen, A., Kjems, J., & Andreasen, P. A. (2010). RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1. Biochemistry, 49(19), 4103-15. https://doi.org/10.1021/bi100066j

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MLA

Vancouver

Author

Madsen, Jeppe B ; Dupont, Daniel Miotto ; Andersen, Thomas B ; Nielsen, Anne F ; Sang, Lu ; Brix, Ditte M ; Jensen, Jan K ; Broos, Thomas ; Hendrickx, Maarten L V ; Christensen, Anni ; Kjems, Jørgen ; Andreasen, Peter A. / RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1. I: Biochemistry. 2010 ; Bind 49, Nr. 19. s. 4103-15.

Bibtex

@article{44cee1d4137d424fa0fdcb7b27d647ad,
title = "RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1",
abstract = "The hallmark of serpins is the ability to undergo the so-called {"}stressed-to-relaxed{"} switch during which the surface-exposed reactive center loop (RCL) becomes incorporated as strand 4 in central beta-sheet A. RCL insertion drives not only the inhibitory reaction of serpins with their target serine proteases but also the conversion to the inactive latent state. RCL insertion is coupled to conformational changes in the flexible joint region flanking beta-sheet A. One interesting serpin is plasminogen activator inhibitor-1 (PAI-1), a fast and specific inhibitor of the serine proteases tissue-type and urokinase-type plasminogen activator. Via its flexible joints' region, native PAI-1 binds vitronectin and relaxed, protease-complexed PAI-1 certain endocytosis receptors. From a library of 35-nucleotides long 2'-fluoropyrimidine-containing RNA oligonucleotides, we have isolated two aptamers binding PAI-1 by the flexible joint region with low nanomolar K(D) values. One of the aptamers exhibited measurable binding to native PAI-1 only, while the other also bound relaxed PAI-1. While none of the aptamers inhibited the antiproteolytic effect of PAI-1, both aptamers inhibited vitronectin binding and the relaxed PAI-1-binding aptamer also endocytosis receptor binding. The aptamer binding exclusively to native PAI-1 increased the half-life for the latency transition to more than 6 h, manyfold more than vitronectin. Contact with Lys124 in the flexible joint region was critical for strong inhibition of the latency transition and the lack of binding to relaxed PAI-1. We conclude that aptamers yield important information about the serpin conformational switch and, because they can compete with high-affinity protein-protein interactions, may provide leads for pharmacological intervention.",
keywords = "Aptamers, Nucleotide, Base Sequence, Binding Sites, Crystallography, X-Ray, Humans, Kinetics, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Plasminogen Activator Inhibitor 1, Protein Conformation",
author = "Madsen, {Jeppe B} and Dupont, {Daniel Miotto} and Andersen, {Thomas B} and Nielsen, {Anne F} and Lu Sang and Brix, {Ditte M} and Jensen, {Jan K} and Thomas Broos and Hendrickx, {Maarten L V} and Anni Christensen and J{\o}rgen Kjems and Andreasen, {Peter A}",
year = "2010",
month = may,
day = "18",
doi = "10.1021/bi100066j",
language = "English",
volume = "49",
pages = "4103--15",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "ACS Publications",
number = "19",

}

RIS

TY - JOUR

T1 - RNA aptamers as conformational probes and regulatory agents for plasminogen activator inhibitor-1

AU - Madsen, Jeppe B

AU - Dupont, Daniel Miotto

AU - Andersen, Thomas B

AU - Nielsen, Anne F

AU - Sang, Lu

AU - Brix, Ditte M

AU - Jensen, Jan K

AU - Broos, Thomas

AU - Hendrickx, Maarten L V

AU - Christensen, Anni

AU - Kjems, Jørgen

AU - Andreasen, Peter A

PY - 2010/5/18

Y1 - 2010/5/18

N2 - The hallmark of serpins is the ability to undergo the so-called "stressed-to-relaxed" switch during which the surface-exposed reactive center loop (RCL) becomes incorporated as strand 4 in central beta-sheet A. RCL insertion drives not only the inhibitory reaction of serpins with their target serine proteases but also the conversion to the inactive latent state. RCL insertion is coupled to conformational changes in the flexible joint region flanking beta-sheet A. One interesting serpin is plasminogen activator inhibitor-1 (PAI-1), a fast and specific inhibitor of the serine proteases tissue-type and urokinase-type plasminogen activator. Via its flexible joints' region, native PAI-1 binds vitronectin and relaxed, protease-complexed PAI-1 certain endocytosis receptors. From a library of 35-nucleotides long 2'-fluoropyrimidine-containing RNA oligonucleotides, we have isolated two aptamers binding PAI-1 by the flexible joint region with low nanomolar K(D) values. One of the aptamers exhibited measurable binding to native PAI-1 only, while the other also bound relaxed PAI-1. While none of the aptamers inhibited the antiproteolytic effect of PAI-1, both aptamers inhibited vitronectin binding and the relaxed PAI-1-binding aptamer also endocytosis receptor binding. The aptamer binding exclusively to native PAI-1 increased the half-life for the latency transition to more than 6 h, manyfold more than vitronectin. Contact with Lys124 in the flexible joint region was critical for strong inhibition of the latency transition and the lack of binding to relaxed PAI-1. We conclude that aptamers yield important information about the serpin conformational switch and, because they can compete with high-affinity protein-protein interactions, may provide leads for pharmacological intervention.

AB - The hallmark of serpins is the ability to undergo the so-called "stressed-to-relaxed" switch during which the surface-exposed reactive center loop (RCL) becomes incorporated as strand 4 in central beta-sheet A. RCL insertion drives not only the inhibitory reaction of serpins with their target serine proteases but also the conversion to the inactive latent state. RCL insertion is coupled to conformational changes in the flexible joint region flanking beta-sheet A. One interesting serpin is plasminogen activator inhibitor-1 (PAI-1), a fast and specific inhibitor of the serine proteases tissue-type and urokinase-type plasminogen activator. Via its flexible joints' region, native PAI-1 binds vitronectin and relaxed, protease-complexed PAI-1 certain endocytosis receptors. From a library of 35-nucleotides long 2'-fluoropyrimidine-containing RNA oligonucleotides, we have isolated two aptamers binding PAI-1 by the flexible joint region with low nanomolar K(D) values. One of the aptamers exhibited measurable binding to native PAI-1 only, while the other also bound relaxed PAI-1. While none of the aptamers inhibited the antiproteolytic effect of PAI-1, both aptamers inhibited vitronectin binding and the relaxed PAI-1-binding aptamer also endocytosis receptor binding. The aptamer binding exclusively to native PAI-1 increased the half-life for the latency transition to more than 6 h, manyfold more than vitronectin. Contact with Lys124 in the flexible joint region was critical for strong inhibition of the latency transition and the lack of binding to relaxed PAI-1. We conclude that aptamers yield important information about the serpin conformational switch and, because they can compete with high-affinity protein-protein interactions, may provide leads for pharmacological intervention.

KW - Aptamers, Nucleotide

KW - Base Sequence

KW - Binding Sites

KW - Crystallography, X-Ray

KW - Humans

KW - Kinetics

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Nucleic Acid Conformation

KW - Plasminogen Activator Inhibitor 1

KW - Protein Conformation

U2 - 10.1021/bi100066j

DO - 10.1021/bi100066j

M3 - Journal article

C2 - 20387790

VL - 49

SP - 4103

EP - 4115

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 19

ER -