SrasV1, Main, Exploration, bibRecord, 005041

Synthesis and Characterization of a Native, Oligomeric Form of Recombinant Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein

Identifieur interne : 005041 ( Main/Exploration ); précédent : 005040; suivant : 005042

Synthesis and Characterization of a Native, Oligomeric Form of Recombinant Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein

Auteurs : Hyun Chul Song ; Mi-Young Seo ; Konrad Stadler ; Byoung J. Yoo ; Qui-Lim Choo ; Stephen R. Coates ; Yasushi Uematsu ; Takashi Harada ; Catherine E. Greer ; John M. Polo ; Piero Pileri ; Markus Eickmann ; Rino Rappuoli ; Sergio Abrignani ; Michael Houghton ; Jang H. Han

Source :

Journal of Virology [ 0022-538X ] ; 2004.

RBID : PMC:516425

Descripteurs français

English descriptors

KwdEn :
- Animals, Antigens, Viral (chemistry), Antigens, Viral (genetics), Antigens, Viral (immunology), Antigens, Viral (metabolism), COS Cells, Cell Line, Chlorocebus aethiops, Cricetinae, Culture Media (chemistry), DNA, Complementary, DNA, Viral (genetics), DNA, Viral (metabolism), Endoplasmic Reticulum (chemistry), Glycoside Hydrolases (metabolism), Golgi Apparatus (chemistry), Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Membrane Glycoproteins (metabolism), Molecular Weight, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase (metabolism), Protein Folding, Protein Processing, Post-Translational, Protein Structure, Tertiary, Protein Subunits (analysis), Protein Transport, Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (metabolism), SARS Virus (genetics), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology), Viral Envelope Proteins (metabolism).
MESH :
- chemical , analysis : Protein Subunits.
- chemical , chemistry : Antigens, Viral, Culture Media, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , genetics : Antigens, Viral, DNA, Viral, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , immunology : Antigens, Viral, Membrane Glycoproteins, Recombinant Proteins, Viral Envelope Proteins.
- chemical , metabolism : Antigens, Viral, DNA, Viral, Glycoside Hydrolases, Membrane Glycoproteins, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, Recombinant Proteins, Viral Envelope Proteins.
- chemistry : Endoplasmic Reticulum, Golgi Apparatus.
- genetics : SARS Virus.
- Animals, COS Cells, Cell Line, Chlorocebus aethiops, Cricetinae, DNA, Complementary, Molecular Weight, Protein Folding, Protein Processing, Post-Translational, Protein Structure, Tertiary, Protein Transport, Spike Glycoprotein, Coronavirus.

Abstract

We have expressed and characterized the severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein in cDNA-transfected mammalian cells. The full-length spike protein (S) was newly synthesized as an endoglycosidase H (endo H)-sensitive glycoprotein (gp170) that is further modified into an endo H-resistant glycoprotein (gp180) in the Golgi apparatus. No substantial proteolytic cleavage of S was observed, suggesting that S is not processed into head (S1) and stalk (S2) domains as observed for certain other coronaviruses. While the expressed full-length S glycoprotein was exclusively cell associated, a truncation of S by excluding the C-terminal transmembrane and cytoplasmic tail domains resulted in the expression of an endoplasmic reticulum-localized glycoprotein (gp160) as well as a Golgi-specific form (gp170) which was ultimately secreted into the cell culture medium. Chemical cross-linking, thermal denaturation, and size fractionation analyses suggested that the full-length S glycoprotein of SARS-CoV forms a higher order structure of ∼500 kDa, which is consistent with it being an S homotrimer. The latter was also observed in purified virions. The intracellular form of the C-terminally truncated S protein (but not the secreted form) also forms trimers, but with much less efficiency than full-length S. Deglycosylation of the full-length homotrimer with peptide N-glycosidase-F under native conditions abolished recognition of the protein by virus-neutralizing antisera raised against purified virions, suggesting the importance of the carbohydrate in the correct folding of the S protein. These data should aid in the design of recombinant vaccine antigens to prevent the spread of this emerging pathogen.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC516425

DOI: 10.1128/JVI.78.19.10328-10335.2004
PubMed: 15367599
PubMed Central: 516425

Affiliations:

Le document en format XML

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<term>Antigens, Viral (immunology)</term>
<term>Antigens, Viral (metabolism)</term>
<term>COS Cells</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
<term>Cricetinae</term>
<term>Culture Media (chemistry)</term>
<term>DNA, Complementary</term>
<term>DNA, Viral (genetics)</term>
<term>DNA, Viral (metabolism)</term>
<term>Endoplasmic Reticulum (chemistry)</term>
<term>Glycoside Hydrolases (metabolism)</term>
<term>Golgi Apparatus (chemistry)</term>
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<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Membrane Glycoproteins (metabolism)</term>
<term>Molecular Weight</term>
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<term>COS Cells</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
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<front><div type="abstract" xml:lang="en"><p>We have expressed and characterized the severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein in cDNA-transfected mammalian cells. The full-length spike protein (S) was newly synthesized as an endoglycosidase H (endo H)-sensitive glycoprotein (gp170) that is further modified into an endo H-resistant glycoprotein (gp180) in the Golgi apparatus. No substantial proteolytic cleavage of S was observed, suggesting that S is not processed into head (S1) and stalk (S2) domains as observed for certain other coronaviruses. While the expressed full-length S glycoprotein was exclusively cell associated, a truncation of S by excluding the C-terminal transmembrane and cytoplasmic tail domains resulted in the expression of an endoplasmic reticulum-localized glycoprotein (gp160) as well as a Golgi-specific form (gp170) which was ultimately secreted into the cell culture medium. Chemical cross-linking, thermal denaturation, and size fractionation analyses suggested that the full-length S glycoprotein of SARS-CoV forms a higher order structure of ∼500 kDa, which is consistent with it being an S homotrimer. The latter was also observed in purified virions. The intracellular form of the C-terminally truncated S protein (but not the secreted form) also forms trimers, but with much less efficiency than full-length S. Deglycosylation of the full-length homotrimer with peptide <italic>N-</italic>
glycosidase-F under native conditions abolished recognition of the protein by virus-neutralizing antisera raised against purified virions, suggesting the importance of the carbohydrate in the correct folding of the S protein. These data should aid in the design of recombinant vaccine antigens to prevent the spread of this emerging pathogen.</p>
</div>
</front>
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<tree><noCountry><name sortKey="Abrignani, Sergio" sort="Abrignani, Sergio" uniqKey="Abrignani S" first="Sergio" last="Abrignani">Sergio Abrignani</name>
<name sortKey="Choo, Qui Lim" sort="Choo, Qui Lim" uniqKey="Choo Q" first="Qui-Lim" last="Choo">Qui-Lim Choo</name>
<name sortKey="Coates, Stephen R" sort="Coates, Stephen R" uniqKey="Coates S" first="Stephen R." last="Coates">Stephen R. Coates</name>
<name sortKey="Eickmann, Markus" sort="Eickmann, Markus" uniqKey="Eickmann M" first="Markus" last="Eickmann">Markus Eickmann</name>
<name sortKey="Greer, Catherine E" sort="Greer, Catherine E" uniqKey="Greer C" first="Catherine E." last="Greer">Catherine E. Greer</name>
<name sortKey="Han, Jang H" sort="Han, Jang H" uniqKey="Han J" first="Jang H." last="Han">Jang H. Han</name>
<name sortKey="Harada, Takashi" sort="Harada, Takashi" uniqKey="Harada T" first="Takashi" last="Harada">Takashi Harada</name>
<name sortKey="Houghton, Michael" sort="Houghton, Michael" uniqKey="Houghton M" first="Michael" last="Houghton">Michael Houghton</name>
<name sortKey="Pileri, Piero" sort="Pileri, Piero" uniqKey="Pileri P" first="Piero" last="Pileri">Piero Pileri</name>
<name sortKey="Polo, John M" sort="Polo, John M" uniqKey="Polo J" first="John M." last="Polo">John M. Polo</name>
<name sortKey="Rappuoli, Rino" sort="Rappuoli, Rino" uniqKey="Rappuoli R" first="Rino" last="Rappuoli">Rino Rappuoli</name>
<name sortKey="Seo, Mi Young" sort="Seo, Mi Young" uniqKey="Seo M" first="Mi-Young" last="Seo">Mi-Young Seo</name>
<name sortKey="Song, Hyun Chul" sort="Song, Hyun Chul" uniqKey="Song H" first="Hyun Chul" last="Song">Hyun Chul Song</name>
<name sortKey="Stadler, Konrad" sort="Stadler, Konrad" uniqKey="Stadler K" first="Konrad" last="Stadler">Konrad Stadler</name>
<name sortKey="Uematsu, Yasushi" sort="Uematsu, Yasushi" uniqKey="Uematsu Y" first="Yasushi" last="Uematsu">Yasushi Uematsu</name>
<name sortKey="Yoo, Byoung J" sort="Yoo, Byoung J" uniqKey="Yoo B" first="Byoung J." last="Yoo">Byoung J. Yoo</name>
</noCountry>
</tree>
</affiliations>
</record>

Pour manipuler ce document sous Unix (Dilib)

EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration

HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 005041 | SxmlIndent | more

HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 005041 | SxmlIndent | more

Pour mettre un lien sur cette page dans le réseau Wicri

{{Explor lien
   |wiki=    Sante
   |area=    SrasV1
   |flux=    Main
   |étape=   Exploration
   |type=    RBID
   |clé=     PMC:516425
   |texte=   Synthesis and Characterization of a Native, Oligomeric Form of Recombinant Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein
}}

Pour générer des pages wiki

HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i   -Sk "pubmed:15367599" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd   \
       | NlmPubMed2Wicri -a SrasV1

This area was generated with Dilib version V0.6.33.
Data generation: Tue Apr 28 14:49:16 2020. Site generation: Sat Mar 27 22:06:49 2021

Serveur d'exploration SRAS

Synthesis and Characterization of a Native, Oligomeric Form of Recombinant Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein

Synthesis and Characterization of a Native, Oligomeric Form of Recombinant Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein

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Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

	Serveur d'exploration SRAS
	Attention, ce site est en cours de développement ! Attention, site généré par des moyens informatiques à partir de corpus bruts. Les informations ne sont donc pas validées.