SrasV1, Main, Exploration, bibRecord, 002962

mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein*

Identifieur interne : 002962 ( Main/Exploration ); précédent : 002961; suivant : 002963

mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein*

Auteurs : Hsiang-I. Liao ; C. Anders Olson [États-Unis] ; Seungmin Hwang ; Hongyu Deng ; Elaine Wong ; Ralph S. Baric ; Richard W. Roberts ; Ren Sun

Source :

The Journal of Biological Chemistry [ 0021-9258 ] ; 2009.

RBID : PMC:2719390

Descripteurs français

KwdFr :
- ARN messager (génétique), ARN messager (métabolisme), Analyse de profil d'expression de gènes, Animaux, Anticorps monoclonaux (génétique), Anticorps monoclonaux (immunologie), Banque de peptides, Cellules Vero, Cellules cultivées, Données de séquences moléculaires, Fibronectines (génétique), Fibronectines (immunologie), Fibronectines (métabolisme), Humains, Immunoprécipitation, Protéines nucléocapside (antagonistes et inhibiteurs), Protéines nucléocapside (génétique), Protéines nucléocapside (métabolisme), RT-PCR, Rein (cytologie), Rein (métabolisme), Réplication virale, Similitude de séquences d'acides aminés, Syndrome respiratoire aigu sévère, Séquence d'acides aminés, Technique d'immunofluorescence, Virus du SRAS (physiologie).
MESH :
- antagonistes et inhibiteurs : Protéines nucléocapside.
- cytologie : Rein.
- génétique : ARN messager, Anticorps monoclonaux, Fibronectines, Protéines nucléocapside.
- immunologie : Anticorps monoclonaux, Fibronectines.
- métabolisme : ARN messager, Fibronectines, Protéines nucléocapside, Rein.
- physiologie : Virus du SRAS.
- Analyse de profil d'expression de gènes, Animaux, Banque de peptides, Cellules Vero, Cellules cultivées, Données de séquences moléculaires, Humains, Immunoprécipitation, RT-PCR, Réplication virale, Similitude de séquences d'acides aminés, Syndrome respiratoire aigu sévère, Séquence d'acides aminés, Technique d'immunofluorescence.

English descriptors

KwdEn :
- Amino Acid Sequence, Animals, Antibodies, Monoclonal (genetics), Antibodies, Monoclonal (immunology), Cells, Cultured, Chlorocebus aethiops, Fibronectins (genetics), Fibronectins (immunology), Fibronectins (metabolism), Fluorescent Antibody Technique, Gene Expression Profiling, Humans, Immunoprecipitation, Kidney (cytology), Kidney (metabolism), Molecular Sequence Data, Nucleocapsid Proteins (antagonists & inhibitors), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (metabolism), Peptide Library, RNA, Messenger (genetics), RNA, Messenger (metabolism), Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (physiology), Sequence Homology, Amino Acid, Severe Acute Respiratory Syndrome, Vero Cells, Virus Replication.
MESH :
- chemical , antagonists & inhibitors : Nucleocapsid Proteins.
- chemical , genetics : Antibodies, Monoclonal, Fibronectins, Nucleocapsid Proteins, RNA, Messenger.
- chemical , immunology : Antibodies, Monoclonal, Fibronectins.
- chemical , metabolism : Fibronectins, Nucleocapsid Proteins, RNA, Messenger.
- cytology : Kidney.
- metabolism : Kidney.
- physiology : SARS Virus.
- Amino Acid Sequence, Animals, Cells, Cultured, Chlorocebus aethiops, Fluorescent Antibody Technique, Gene Expression Profiling, Humans, Immunoprecipitation, Molecular Sequence Data, Peptide Library, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Severe Acute Respiratory Syndrome, Vero Cells, Virus Replication.

Abstract

The nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus plays important roles in both viral replication and modulation of host cell processes. New ligands that target the N protein may thus provide tools to track the protein inside cells, detect interaction hot spots on the protein surface, and discover sites that could be used to develop new anti-SARS therapies. Using mRNA display selection and directed evolution, we designed novel antibody-like protein affinity reagents that target SARS N protein with high affinity and selectivity. Our libraries were based on an 88-residue variant of the 10th fibronectin type III domain from human fibronectin (10Fn3). This selection resulted in eight independent 10Fn3 intrabodies, two that require the N-terminal domain for binding and six that recognize the C terminus, one with K_d = 1.7 nm. 10Fn3 intrabodies are well expressed in mammalian cells and are relocalized by N in SARS-infected cells. Seven of the selected intrabodies tested do not perturb cellular function when expressed singly in vivo and inhibit virus replication from 11- to 5900-fold when expressed in cells prior to infection. Targeting two sites on SARS-N simultaneously using two distinct 10Fn3s results in synergistic inhibition of virus replication.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2719390

DOI: 10.1074/jbc.M901547200
PubMed: 19364769
PubMed Central: 2719390

Affiliations:

Le document en format XML

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<author><name sortKey="Wong, Elaine" sort="Wong, Elaine" uniqKey="Wong E" first="Elaine" last="Wong">Elaine Wong</name>
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<author><name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Animals</term>
<term>Antibodies, Monoclonal (genetics)</term>
<term>Antibodies, Monoclonal (immunology)</term>
<term>Cells, Cultured</term>
<term>Chlorocebus aethiops</term>
<term>Fibronectins (genetics)</term>
<term>Fibronectins (immunology)</term>
<term>Fibronectins (metabolism)</term>
<term>Fluorescent Antibody Technique</term>
<term>Gene Expression Profiling</term>
<term>Humans</term>
<term>Immunoprecipitation</term>
<term>Kidney (cytology)</term>
<term>Kidney (metabolism)</term>
<term>Molecular Sequence Data</term>
<term>Nucleocapsid Proteins (antagonists & inhibitors)</term>
<term>Nucleocapsid Proteins (genetics)</term>
<term>Nucleocapsid Proteins (metabolism)</term>
<term>Peptide Library</term>
<term>RNA, Messenger (genetics)</term>
<term>RNA, Messenger (metabolism)</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>SARS Virus (physiology)</term>
<term>Sequence Homology, Amino Acid</term>
<term>Severe Acute Respiratory Syndrome</term>
<term>Vero Cells</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN messager (génétique)</term>
<term>ARN messager (métabolisme)</term>
<term>Analyse de profil d'expression de gènes</term>
<term>Animaux</term>
<term>Anticorps monoclonaux (génétique)</term>
<term>Anticorps monoclonaux (immunologie)</term>
<term>Banque de peptides</term>
<term>Cellules Vero</term>
<term>Cellules cultivées</term>
<term>Données de séquences moléculaires</term>
<term>Fibronectines (génétique)</term>
<term>Fibronectines (immunologie)</term>
<term>Fibronectines (métabolisme)</term>
<term>Humains</term>
<term>Immunoprécipitation</term>
<term>Protéines nucléocapside (antagonistes et inhibiteurs)</term>
<term>Protéines nucléocapside (génétique)</term>
<term>Protéines nucléocapside (métabolisme)</term>
<term>RT-PCR</term>
<term>Rein (cytologie)</term>
<term>Rein (métabolisme)</term>
<term>Réplication virale</term>
<term>Similitude de séquences d'acides aminés</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Séquence d'acides aminés</term>
<term>Technique d'immunofluorescence</term>
<term>Virus du SRAS (physiologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>Nucleocapsid Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antibodies, Monoclonal</term>
<term>Fibronectins</term>
<term>Nucleocapsid Proteins</term>
<term>RNA, Messenger</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term>
<term>Fibronectins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Fibronectins</term>
<term>Nucleocapsid Proteins</term>
<term>RNA, Messenger</term>
</keywords>
<keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Rein</term>
</keywords>
<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Kidney</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN messager</term>
<term>Anticorps monoclonaux</term>
<term>Fibronectines</term>
<term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps monoclonaux</term>
<term>Fibronectines</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Kidney</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN messager</term>
<term>Fibronectines</term>
<term>Protéines nucléocapside</term>
<term>Rein</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Animals</term>
<term>Cells, Cultured</term>
<term>Chlorocebus aethiops</term>
<term>Fluorescent Antibody Technique</term>
<term>Gene Expression Profiling</term>
<term>Humans</term>
<term>Immunoprecipitation</term>
<term>Molecular Sequence Data</term>
<term>Peptide Library</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>Sequence Homology, Amino Acid</term>
<term>Severe Acute Respiratory Syndrome</term>
<term>Vero Cells</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Analyse de profil d'expression de gènes</term>
<term>Animaux</term>
<term>Banque de peptides</term>
<term>Cellules Vero</term>
<term>Cellules cultivées</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Immunoprécipitation</term>
<term>RT-PCR</term>
<term>Réplication virale</term>
<term>Similitude de séquences d'acides aminés</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Séquence d'acides aminés</term>
<term>Technique d'immunofluorescence</term>
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<front><div type="abstract" xml:lang="en"><p>The nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus plays important roles in both viral replication and modulation of host cell processes. New ligands that target the N protein may thus provide tools to track the protein inside cells, detect interaction hot spots on the protein surface, and discover sites that could be used to develop new anti-SARS therapies. Using mRNA display selection and directed evolution, we designed novel antibody-like protein affinity reagents that target SARS N protein with high affinity and selectivity. Our libraries were based on an 88-residue variant of the 10th fibronectin type III domain from human fibronectin (10Fn3). This selection resulted in eight independent 10Fn3 intrabodies, two that require the N-terminal domain for binding and six that recognize the C terminus, one with <italic>K<sub>d</sub>
</italic>
 = 1.7 n<sc>m</sc>
. 10Fn3 intrabodies are well expressed in mammalian cells and are relocalized by N in SARS-infected cells. Seven of the selected intrabodies tested do not perturb cellular function when expressed singly <italic>in vivo</italic>
 and inhibit virus replication from 11- to 5900-fold when expressed in cells prior to infection. Targeting two sites on SARS-N simultaneously using two distinct 10Fn3s results in synergistic inhibition of virus replication.</p>
</div>
</front>
</TEI>
<affiliations><list><country><li>États-Unis</li>
</country>
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</region>
</list>
<tree><noCountry><name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name>
<name sortKey="Deng, Hongyu" sort="Deng, Hongyu" uniqKey="Deng H" first="Hongyu" last="Deng">Hongyu Deng</name>
<name sortKey="Hwang, Seungmin" sort="Hwang, Seungmin" uniqKey="Hwang S" first="Seungmin" last="Hwang">Seungmin Hwang</name>
<name sortKey="Liao, Hsiang I" sort="Liao, Hsiang I" uniqKey="Liao H" first="Hsiang-I." last="Liao">Hsiang-I. Liao</name>
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<name sortKey="Sun, Ren" sort="Sun, Ren" uniqKey="Sun R" first="Ren" last="Sun">Ren Sun</name>
<name sortKey="Wong, Elaine" sort="Wong, Elaine" uniqKey="Wong E" first="Elaine" last="Wong">Elaine Wong</name>
</noCountry>
<country name="États-Unis"><region name="Californie"><name sortKey="Olson, C Anders" sort="Olson, C Anders" uniqKey="Olson C" first="C. Anders" last="Olson">C. Anders Olson</name>
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mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein*

mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein*

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

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