期刊文献+

Dissection of SARS Coronavirus Spike Protein into Discrete Folded Fragments

Dissection of SARS Coronavirus Spike Protein into Discrete Folded Fragments
原文传递
导出
摘要 The spike protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) mediates cell fusion by binding to target cell surface receptors. This paper reports a simple method for dissecting the viral protein and for searching for foldable fragments in a random but systematic manner. The method involves digestion by DNase I to generate a pool of short DNA segments, followed by an additional step of reassembly of these segments to produce a library of DNA fragments with random ends but controllable lengths. To rapidly screen for discrete folded polypeptide fragments, the reassembled gene fragments were further cloned into a vector as N-terminal fusions to a folding reporter gene which was a variant of green fluorescent protein. Two foldable fragments were identified for the SARS-CoV spike protein, which coincide with various anti-SARS peptides derived from the hepated repeat (HR) region 2 of the spike protein. The method should be applicable to other viral proteins to isolate antigen or vaccine candidates, thus providing an alternative to the full-length proteins (subunits) or linear short peptides. The spike protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) mediates cell fusion by binding to target cell surface receptors. This paper reports a simple method for dissecting the viral protein and for searching for foldable fragments in a random but systematic manner. The method involves digestion by DNase I to generate a pool of short DNA segments, followed by an additional step of reassembly of these segments to produce a library of DNA fragments with random ends but controllable lengths. To rapidly screen for discrete folded polypeptide fragments, the reassembled gene fragments were further cloned into a vector as N-terminal fusions to a folding reporter gene which was a variant of green fluorescent protein. Two foldable fragments were identified for the SARS-CoV spike protein, which coincide with various anti-SARS peptides derived from the hepated repeat (HR) region 2 of the spike protein. The method should be applicable to other viral proteins to isolate antigen or vaccine candidates, thus providing an alternative to the full-length proteins (subunits) or linear short peptides.
出处 《Tsinghua Science and Technology》 SCIE EI CAS 2006年第4期490-494,共5页 清华大学学报(自然科学版(英文版)
基金 Supported by the Tsinghua University SARS Special Fund and theNational Key Basic Research and Development (973) Program of China (No. 2003CB716002)
关键词 severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein DISSECTION foldablefragment green fluorscent protein (GFP) severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein dissection foldablefragment green fluorscent protein (GFP)
  • 相关文献

参考文献19

  • 1Harbury P;Zhang T;Kim P.A switch between two-,three-,and four-stranded coiled coils in GCN4 leucine zipper mutants[J],1993(5138).
  • 2Gaboriaud C;Bissery V;Benchetrit T.Hydrophobic cluster analysis:An efficient new way to compare and analyze amino acid sequences[J],1987(01).
  • 3Tripet B;Howard M W;Jobling M.Structural characterization of the SARS-coronavirus spike (S) fusion protein core[J],2004(20).
  • 4Kimata Y;Iwaki M;Lim C R.A novel mutation which enhances the fluorescence of green fluorescent protein at high temperatures[J],1997(01).
  • 5Zhu J Q;Xiao G F;Xu Y H.Following the rule:Formation of the 6-helix bundle of the fusion core from severe acute respiratory syndrome coronavirus spike protein and identification of potent peptide inhibitors[J],2004(01).
  • 6Liu S W;Xiao G F;Chen Y B.Interaction between heptad repeat 1 and 2 regions in spike protein of SARS-associated coronavirus:Implications for virus fusogenic mechanism and identification of fusion inhibitors[J],2004(9413).
  • 7Bosch B J;Martina B E E;van der Zee R.Severe acute respiratory syndrome coroavirus (SARS-CoV) infection inhibition using spike protein heptad repeat-derived peptides[J],2004(22).
  • 8Waldo G S.Genetic screens and directed evolution for protein solubility[J],2003(01).
  • 9Waldo G S;Standish B M;Berendzen J.Rapid protein-folding assay using green fluorescent protein[J],1999(07).
  • 10Holmes K V;Zelus B D;Schickli J H.Receptor specificity and receptor-induced conformational changes in mouse hepatitis virus spike glycoprotein,2001.

共引文献7

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部