دورية أكاديمية
Synthesis and characterization of a native, oligomeric form of recombinant severe acute respiratory syndrome coronavirus spike glycoprotein.
| العنوان: | Synthesis and characterization of a native, oligomeric form of recombinant severe acute respiratory syndrome coronavirus spike glycoprotein. |
|---|---|
| المؤلفون: | Song HC; Vaccines Research, Chiron Corporation, 4560 Horton St., Emeryville, CA 94608, USA., Seo MY, Stadler K, Yoo BJ, Choo QL, Coates SR, Uematsu Y, Harada T, Greer CE, Polo JM, Pileri P, Eickmann M, Rappuoli R, Abrignani S, Houghton M, Han JH |
| المصدر: | Journal of virology [J Virol] 2004 Oct; Vol. 78 (19), pp. 10328-35. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society For Microbiology Country of Publication: United States NLM ID: 0113724 Publication Model: Print Cited Medium: Print ISSN: 0022-538X (Print) Linking ISSN: 0022538X NLM ISO Abbreviation: J Virol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Washington Dc : American Society For Microbiology Original Publication: Baltimore, American Society for Microbiology. |
| مواضيع طبية MeSH: | Membrane Glycoproteins/*chemistry , Membrane Glycoproteins/*metabolism , Severe acute respiratory syndrome-related coronavirus/*genetics , Viral Envelope Proteins/*chemistry , Viral Envelope Proteins/*metabolism, 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/genetics ; Membrane Glycoproteins/immunology ; 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 ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins/genetics ; Viral Envelope Proteins/immunology |
| مستخلص: | 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. |
| References: | Virology. 2000 Mar 30;269(1):212-24. (PMID: 10725213) J Virol. 1999 Oct;73(10):8152-9. (PMID: 10482565) N Engl J Med. 2003 May 15;348(20):1953-66. (PMID: 12690092) Science. 2003 May 30;300(5624):1394-9. (PMID: 12730500) Science. 2003 May 30;300(5624):1399-404. (PMID: 12730501) Science. 2003 May 30;300(5624):1351. (PMID: 12775803) J Virol. 2003 Aug;77(16):8801-11. (PMID: 12885899) J Virol. 2003 Oct;77(19):10394-403. (PMID: 12970424) J Virol. 2003 Oct;77(20):11244-59. (PMID: 14512572) Science. 2003 Oct 10;302(5643):276-8. (PMID: 12958366) Nature. 2003 Oct 30;425(6961):915. (PMID: 14586458) Biochem Biophys Res Commun. 2003 Dec 26;312(4):1159-64. (PMID: 14651994) Nature. 2003 Nov 27;426(6965):450-4. (PMID: 14647384) Science. 2003 Nov 28;302(5650):1504-5. (PMID: 14645828) Avian Pathol. 2003 Dec;32(6):567-82. (PMID: 14676007) J Biol Chem. 2004 Jan 30;279(5):3197-201. (PMID: 14670965) BMC Microbiol. 2003 Sep 21;3:20. (PMID: 14499001) Biochem Biophys Res Commun. 2004 Mar 5;315(2):439-44. (PMID: 14766227) Virology. 1977 Jun 15;79(2):446-8. (PMID: 867833) Nature. 1981 Jan 29;289(5796):366-73. (PMID: 7464906) Cell. 1986 Sep 12;46(6):929-37. (PMID: 3019557) Cell. 1986 Sep 12;46(6):939-50. (PMID: 3757030) J Cell Biol. 1986 Oct;103(4):1179-91. (PMID: 2429970) J Virol. 1986 Dec;60(3):833-9. (PMID: 3783818) J Cell Biol. 1987 Nov;105(5):1957-69. (PMID: 2824524) Cell. 1988 Mar 25;52(6):843-52. (PMID: 2450677) J Virol. 1990 Nov;64(11):5367-75. (PMID: 2170676) Virology. 1991 Jun;182(2):765-73. (PMID: 1850927) J Virol. 1993 Nov;67(11):6753-61. (PMID: 8411378) Nature. 1998 Jun 18;393(6686):705-11. (PMID: 9641684) Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4598-603. (PMID: 10200308) J Virol. 2002 Dec;76(24):12491-502. (PMID: 12438575) |
| المشرفين على المادة: | 0 (Antigens, Viral) 0 (Culture Media) 0 (DNA, Complementary) 0 (DNA, Viral) 0 (Membrane Glycoproteins) 0 (Protein Subunits) 0 (Recombinant Proteins) 0 (Spike Glycoprotein, Coronavirus) 0 (Viral Envelope Proteins) 0 (spike glycoprotein, SARS-CoV) 0 (spike protein, mouse hepatitis virus) EC 3.2.1.- (Glycoside Hydrolases) EC 3.5.1.52 (Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase) |
| تواريخ الأحداث: | Date Created: 20040916 Date Completed: 20041021 Latest Revision: 20221207 |
| رمز التحديث: | 20240829 |
| مُعرف محوري في PubMed: | PMC516425 |
| DOI: | 10.1128/JVI.78.19.10328-10335.2004 |
| PMID: | 15367599 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0022-538X |
|---|---|
| DOI: | 10.1128/JVI.78.19.10328-10335.2004 |