Biochemical and immunological studies of nucleocapsid proteins of severe acute respiratory syndrome and 229E human coronaviruses
Identifieur interne : 004C61 ( Main/Exploration ); précédent : 004C60; suivant : 004C62Biochemical and immunological studies of nucleocapsid proteins of severe acute respiratory syndrome and 229E human coronaviruses
Auteurs : Tswen-Kei Tang ; Mark P. Wu ; Shui-Tsung Chen ; Ming-Hon Hou ; Ming-Hsiang Hong ; Fu-Ming Pan ; Hui-Ming Yu ; Jenn-Han Chen [Taïwan] ; Chen-Wen Yao [Taïwan] ; Andrew H. Wang [Taïwan]Source :
- PROTEOMICS [ 1615-9853 ] ; 2005-03.
Descripteurs français
- KwdFr :
- ADN (), ADN complémentaire (métabolisme), ARN (), Acides aminés (), Analyse par réseau de protéines (), Animaux, Anticorps antiviraux (), Antigènes (), Antigènes viraux (), Cadres ouverts de lecture, Cellules Vero, Chromatographie sur gel, Clonage moléculaire, Coronavirus humain 229E (métabolisme), Dichroïsme circulaire, Dimérisation, Données de séquences moléculaires, Humains, Lapins, Liaison aux protéines, Microscopie de fluorescence, Nucléocapside (), Peptides (), Protéines nucléocapside (), Protéomique (), Réactifs réticulants (pharmacologie), Similitude de séquences d'acides aminés, Sites de fixation, Structure tertiaire des protéines, Syndrome respiratoire aigu sévère (diagnostic), Séquence d'acides aminés, Test ELISA, Virus du SRAS (métabolisme), Électrophorèse sur gel de polyacrylamide, Épitopes ().
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- métabolisme : ADN complémentaire, Coronavirus humain 229E, Virus du SRAS.
- pharmacologie : Réactifs réticulants.
- ADN, ARN, Acides aminés, Analyse par réseau de protéines, Animaux, Anticorps antiviraux, Antigènes, Antigènes viraux, Cadres ouverts de lecture, Cellules Vero, Chromatographie sur gel, Clonage moléculaire, Dichroïsme circulaire, Dimérisation, Données de séquences moléculaires, Humains, Lapins, Liaison aux protéines, Microscopie de fluorescence, Nucléocapside, Peptides, Protéines nucléocapside, Protéomique, Similitude de séquences d'acides aminés, Sites de fixation, Structure tertiaire des protéines, Séquence d'acides aminés, Test ELISA, Électrophorèse sur gel de polyacrylamide, Épitopes.
English descriptors
- KwdEn :
- Amino Acid Sequence, Amino Acids (chemistry), Animals, Antibodies, Viral (chemistry), Antigens (chemistry), Antigens, Viral (chemistry), Binding Sites, Chlorocebus aethiops, Chromatography, Gel, Circular Dichroism, Cloning, Molecular, Coronavirus 229E, Human (metabolism), Cross-Linking Reagents (pharmacology), DNA (chemistry), DNA, Complementary (metabolism), Dimerization, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Epitopes (chemistry), Humans, Microscopy, Fluorescence, Molecular Sequence Data, Nucleocapsid (chemistry), Nucleocapsid Proteins (chemistry), Open Reading Frames, Peptides (chemistry), Protein Array Analysis (methods), Protein Binding, Protein Structure, Tertiary, Proteomics (methods), RNA (chemistry), Rabbits, SARS Virus (metabolism), Sequence Homology, Amino Acid, Severe Acute Respiratory Syndrome (diagnosis), Vero Cells.
- MESH :
- chemical , chemistry : Amino Acids, Antibodies, Viral, Antigens, Antigens, Viral, DNA, Epitopes, Nucleocapsid Proteins, Peptides, RNA.
- chemistry : Nucleocapsid.
- diagnosis : Severe Acute Respiratory Syndrome.
- metabolism : Coronavirus 229E, Human, DNA, Complementary, SARS Virus.
- methods : Protein Array Analysis, Proteomics.
- chemical , pharmacology : Cross-Linking Reagents.
- Teeft :
- Amino, Amino Acid Sequence, Amino acid, Animals, Antibody, Antigeneicity, Assay, Band shift assay, Binding Sites, Chlorocebus aethiops, Chromatography, Gel, Circular Dichroism, Clin, Cloning, Molecular, Complete sequence, Coronavirus, Coronaviruses, Dimer, Dimer form, Dimerization, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Filtration, Final concentration, Genome, Gmbh, Hcov, Human coronavirus, Humans, Immunization, Immunological, Immunological studies, Kgaa, Major band, Major forms, Microscopy, Fluorescence, Minor bands, Molecular Sequence Data, Nacl, Nucleocapsid, Open Reading Frames, Peptide, Polyclonal, Polyclonal antibodies, Pondr, Pondr analysis, Pondr program, Protein, Protein Binding, Protein Structure, Tertiary, Proteomics, Rabbits, Recombinant, Recombinant sars, Respiratory syndrome, Retention volume, Retention volume graph, Room temperature, Sars, Sars hcov, Sars patients, Sars virus, Sequence Homology, Amino Acid, Sequence alignment, Structural proteins, Synthetic peptides, Verlag, Verlag gmbh, Vero Cells, Weinheim, Weinheim proteomics, Western blot analysis.
Abstract
Severe acute respiratory syndrome (SARS) is a serious health threat and its early diagnosis is important for infection control and potential treatment of the disease. Diagnostic tools require rapid and accurate methods, of which a capture ELISA method may be useful. Toward this goal, we have prepared and characterized soluble full‐length nucleocapsid proteins (N protein) from SARS and 229E human coronaviruses. N proteins form oligomers, mostly as dimers at low concentration. These two N proteins degrade rapidly upon storage and the major degraded N protein is the C‐terminal fragment of amino acid (aa) 169–422. Taken together with other data, we suggest that N protein is a two‐domain protein, with the N‐terminal aa 50–150 as the RNA‐binding domain and the C‐terminal aa 169–422 as the dimerization domain. Polyclonal antibodies against the SARS N protein have been produced and the strong binding sites of the anti‐nucleocapsid protein (NP) antibodies produced were mapped to aa 1–20, aa 150–170 and aa 390–410. These sites are generally consistent with those mapped by sera obtained from SARS patients. The SARS anti‐NP antibody was able to clearly detect SARS virus grown in Vero E6 cells and did not cross‐react with the NP from the human coronavirus 229E. We have predicted several antigenic sites (15–20 amino acids) of S, M and N proteins and produced antibodies against those peptides, some of which could be recognized by sera obtained from SARS patients. Antibodies against the NP peptides could detect the cognate N protein clearly. Further refinement of these antibodies, particularly large‐scale production of monoclonal antibodies, could lead to the development of useful diagnostic kits for diseases associated with SARS and other human coronaviruses.
Url:
DOI: 10.1002/pmic.200401204
Affiliations:
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Le document en format XML
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<term>Amino Acids (chemistry)</term>
<term>Animals</term>
<term>Antibodies, Viral (chemistry)</term>
<term>Antigens (chemistry)</term>
<term>Antigens, Viral (chemistry)</term>
<term>Binding Sites</term>
<term>Chlorocebus aethiops</term>
<term>Chromatography, Gel</term>
<term>Circular Dichroism</term>
<term>Cloning, Molecular</term>
<term>Coronavirus 229E, Human (metabolism)</term>
<term>Cross-Linking Reagents (pharmacology)</term>
<term>DNA (chemistry)</term>
<term>DNA, Complementary (metabolism)</term>
<term>Dimerization</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Epitopes (chemistry)</term>
<term>Humans</term>
<term>Microscopy, Fluorescence</term>
<term>Molecular Sequence Data</term>
<term>Nucleocapsid (chemistry)</term>
<term>Nucleocapsid Proteins (chemistry)</term>
<term>Open Reading Frames</term>
<term>Peptides (chemistry)</term>
<term>Protein Array Analysis (methods)</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
<term>Proteomics (methods)</term>
<term>RNA (chemistry)</term>
<term>Rabbits</term>
<term>SARS Virus (metabolism)</term>
<term>Sequence Homology, Amino Acid</term>
<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
<term>Vero Cells</term>
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<term>ADN complémentaire (métabolisme)</term>
<term>ARN ()</term>
<term>Acides aminés ()</term>
<term>Analyse par réseau de protéines ()</term>
<term>Animaux</term>
<term>Anticorps antiviraux ()</term>
<term>Antigènes ()</term>
<term>Antigènes viraux ()</term>
<term>Cadres ouverts de lecture</term>
<term>Cellules Vero</term>
<term>Chromatographie sur gel</term>
<term>Clonage moléculaire</term>
<term>Coronavirus humain 229E (métabolisme)</term>
<term>Dichroïsme circulaire</term>
<term>Dimérisation</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lapins</term>
<term>Liaison aux protéines</term>
<term>Microscopie de fluorescence</term>
<term>Nucléocapside ()</term>
<term>Peptides ()</term>
<term>Protéines nucléocapside ()</term>
<term>Protéomique ()</term>
<term>Réactifs réticulants (pharmacologie)</term>
<term>Similitude de séquences d'acides aminés</term>
<term>Sites de fixation</term>
<term>Structure tertiaire des protéines</term>
<term>Syndrome respiratoire aigu sévère (diagnostic)</term>
<term>Séquence d'acides aminés</term>
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<term>Virus du SRAS (métabolisme)</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
<term>Épitopes ()</term>
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<term>Antibodies, Viral</term>
<term>Antigens</term>
<term>Antigens, Viral</term>
<term>DNA</term>
<term>Epitopes</term>
<term>Nucleocapsid Proteins</term>
<term>Peptides</term>
<term>RNA</term>
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<term>Coronavirus humain 229E</term>
<term>Virus du SRAS</term>
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<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Réactifs réticulants</term>
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<term>Amino Acid Sequence</term>
<term>Amino acid</term>
<term>Animals</term>
<term>Antibody</term>
<term>Antigeneicity</term>
<term>Assay</term>
<term>Band shift assay</term>
<term>Binding Sites</term>
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<term>Chromatography, Gel</term>
<term>Circular Dichroism</term>
<term>Clin</term>
<term>Cloning, Molecular</term>
<term>Complete sequence</term>
<term>Coronavirus</term>
<term>Coronaviruses</term>
<term>Dimer</term>
<term>Dimer form</term>
<term>Dimerization</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Filtration</term>
<term>Final concentration</term>
<term>Genome</term>
<term>Gmbh</term>
<term>Hcov</term>
<term>Human coronavirus</term>
<term>Humans</term>
<term>Immunization</term>
<term>Immunological</term>
<term>Immunological studies</term>
<term>Kgaa</term>
<term>Major band</term>
<term>Major forms</term>
<term>Microscopy, Fluorescence</term>
<term>Minor bands</term>
<term>Molecular Sequence Data</term>
<term>Nacl</term>
<term>Nucleocapsid</term>
<term>Open Reading Frames</term>
<term>Peptide</term>
<term>Polyclonal</term>
<term>Polyclonal antibodies</term>
<term>Pondr</term>
<term>Pondr analysis</term>
<term>Pondr program</term>
<term>Protein</term>
<term>Protein Binding</term>
<term>Protein Structure, Tertiary</term>
<term>Proteomics</term>
<term>Rabbits</term>
<term>Recombinant</term>
<term>Recombinant sars</term>
<term>Respiratory syndrome</term>
<term>Retention volume</term>
<term>Retention volume graph</term>
<term>Room temperature</term>
<term>Sars</term>
<term>Sars hcov</term>
<term>Sars patients</term>
<term>Sars virus</term>
<term>Sequence Homology, Amino Acid</term>
<term>Sequence alignment</term>
<term>Structural proteins</term>
<term>Synthetic peptides</term>
<term>Verlag</term>
<term>Verlag gmbh</term>
<term>Vero Cells</term>
<term>Weinheim</term>
<term>Weinheim proteomics</term>
<term>Western blot analysis</term>
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<term>ARN</term>
<term>Acides aminés</term>
<term>Analyse par réseau de protéines</term>
<term>Animaux</term>
<term>Anticorps antiviraux</term>
<term>Antigènes</term>
<term>Antigènes viraux</term>
<term>Cadres ouverts de lecture</term>
<term>Cellules Vero</term>
<term>Chromatographie sur gel</term>
<term>Clonage moléculaire</term>
<term>Dichroïsme circulaire</term>
<term>Dimérisation</term>
<term>Données de séquences moléculaires</term>
<term>Humains</term>
<term>Lapins</term>
<term>Liaison aux protéines</term>
<term>Microscopie de fluorescence</term>
<term>Nucléocapside</term>
<term>Peptides</term>
<term>Protéines nucléocapside</term>
<term>Protéomique</term>
<term>Similitude de séquences d'acides aminés</term>
<term>Sites de fixation</term>
<term>Structure tertiaire des protéines</term>
<term>Séquence d'acides aminés</term>
<term>Test ELISA</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
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<front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is a serious health threat and its early diagnosis is important for infection control and potential treatment of the disease. Diagnostic tools require rapid and accurate methods, of which a capture ELISA method may be useful. Toward this goal, we have prepared and characterized soluble full‐length nucleocapsid proteins (N protein) from SARS and 229E human coronaviruses. N proteins form oligomers, mostly as dimers at low concentration. These two N proteins degrade rapidly upon storage and the major degraded N protein is the C‐terminal fragment of amino acid (aa) 169–422. Taken together with other data, we suggest that N protein is a two‐domain protein, with the N‐terminal aa 50–150 as the RNA‐binding domain and the C‐terminal aa 169–422 as the dimerization domain. Polyclonal antibodies against the SARS N protein have been produced and the strong binding sites of the anti‐nucleocapsid protein (NP) antibodies produced were mapped to aa 1–20, aa 150–170 and aa 390–410. These sites are generally consistent with those mapped by sera obtained from SARS patients. The SARS anti‐NP antibody was able to clearly detect SARS virus grown in Vero E6 cells and did not cross‐react with the NP from the human coronavirus 229E. We have predicted several antigenic sites (15–20 amino acids) of S, M and N proteins and produced antibodies against those peptides, some of which could be recognized by sera obtained from SARS patients. Antibodies against the NP peptides could detect the cognate N protein clearly. Further refinement of these antibodies, particularly large‐scale production of monoclonal antibodies, could lead to the development of useful diagnostic kits for diseases associated with SARS and other human coronaviruses.</div>
</front>
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<affiliations><list><country><li>Taïwan</li>
</country>
</list>
<tree><noCountry><name sortKey="Chen, Shui Sung" sort="Chen, Shui Sung" uniqKey="Chen S" first="Shui-Tsung" last="Chen">Shui-Tsung Chen</name>
<name sortKey="Hong, Ming Siang" sort="Hong, Ming Siang" uniqKey="Hong M" first="Ming-Hsiang" last="Hong">Ming-Hsiang Hong</name>
<name sortKey="Hou, Ming On" sort="Hou, Ming On" uniqKey="Hou M" first="Ming-Hon" last="Hou">Ming-Hon Hou</name>
<name sortKey="Pan, Fu Ing" sort="Pan, Fu Ing" uniqKey="Pan F" first="Fu-Ming" last="Pan">Fu-Ming Pan</name>
<name sortKey="Tang, Tswen Ei" sort="Tang, Tswen Ei" uniqKey="Tang T" first="Tswen-Kei" last="Tang">Tswen-Kei Tang</name>
<name sortKey="Wu, Mark P" sort="Wu, Mark P" uniqKey="Wu M" first="Mark P." last="Wu">Mark P. Wu</name>
<name sortKey="Yu, Hui Ing" sort="Yu, Hui Ing" uniqKey="Yu H" first="Hui-Ming" last="Yu">Hui-Ming Yu</name>
</noCountry>
<country name="Taïwan"><noRegion><name sortKey="Chen, Jenn An" sort="Chen, Jenn An" uniqKey="Chen J" first="Jenn-Han" last="Chen">Jenn-Han Chen</name>
</noRegion>
<name sortKey="Wang, Andrew H" sort="Wang, Andrew H" uniqKey="Wang A" first="Andrew H." last="Wang">Andrew H. Wang</name>
<name sortKey="Yao, Chen En" sort="Yao, Chen En" uniqKey="Yao C" first="Chen-Wen" last="Yao">Chen-Wen Yao</name>
</country>
</tree>
</affiliations>
</record>
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