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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
19
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pubmed:dateCreated |
2004-9-15
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pubmed:abstractText |
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 approximately 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.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-10200308,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-10482565,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-10725213,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12438575,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12690092,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12730500,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12730501,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12885899,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12958366,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-12970424,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14499001,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14512572,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14586458,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14647384,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14651994,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14670965,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14676007,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-14766227,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-1850927,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-2170676,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-2429970,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-2450677,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-2824524,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-3019557,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-3757030,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-3783818,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-7464906,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-8411378,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-867833,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15367599-9641684
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Viral,
http://linkedlifedata.com/resource/pubmed/chemical/Culture Media,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Viral,
http://linkedlifedata.com/resource/pubmed/chemical/Glycoside Hydrolases,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Glycoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide-N4-(N-acetyl-beta-glucosamin...,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Subunits,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Viral Envelope Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/spike glycoprotein, coronavirus
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0022-538X
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pubmed:author |
pubmed-author:AbrignaniSergioS,
pubmed-author:ChooQui-LimQL,
pubmed-author:CoatesStephen RSR,
pubmed-author:EickmannMarkusM,
pubmed-author:GreerCatherine ECE,
pubmed-author:HanJang HJH,
pubmed-author:HaradaTakashiT,
pubmed-author:HoughtonMichaelM,
pubmed-author:PileriPieroP,
pubmed-author:PoloJohn MJM,
pubmed-author:RappuoliRinoR,
pubmed-author:SeoMi-YoungMY,
pubmed-author:SongHyun ChulHC,
pubmed-author:StadlerKonradK,
pubmed-author:UematsuYasushiY,
pubmed-author:YooByoung JBJ
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pubmed:issnType |
Print
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pubmed:volume |
78
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
10328-35
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:15367599-Animals,
pubmed-meshheading:15367599-Golgi Apparatus,
pubmed-meshheading:15367599-Molecular Weight,
pubmed-meshheading:15367599-Cricetinae,
pubmed-meshheading:15367599-Culture Media,
pubmed-meshheading:15367599-Glycoside Hydrolases,
pubmed-meshheading:15367599-Protein Subunits,
pubmed-meshheading:15367599-DNA, Viral,
pubmed-meshheading:15367599-Endoplasmic Reticulum,
pubmed-meshheading:15367599-Cell Line,
pubmed-meshheading:15367599-Antigens, Viral,
pubmed-meshheading:15367599-Protein Transport,
pubmed-meshheading:15367599-Cercopithecus aethiops,
pubmed-meshheading:15367599-Protein Structure, Tertiary,
pubmed-meshheading:15367599-Protein Processing, Post-Translational,
pubmed-meshheading:15367599-DNA, Complementary,
pubmed-meshheading:15367599-Membrane Glycoproteins,
pubmed-meshheading:15367599-Viral Envelope Proteins,
pubmed-meshheading:15367599-Recombinant Proteins,
pubmed-meshheading:15367599-Protein Folding
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