Epitope mapping of human respiratory syncytial virus 22K transcription antitermination factor: role of N-terminal sequences in protein folding

J Gen Virol. 2005 Apr;86(Pt 4):1103-1107. doi: 10.1099/vir.0.80712-0.

Abstract

The reactivity of a panel of 12 monoclonal antibodies raised against the human respiratory syncytial virus 22 kDa (22K) protein was tested by Western blotting with a set of 22K deletion mutants. The results obtained identified sequences in the C-terminal half of the 22K polypeptide required for integrity of most antibody epitopes, except for epitope 112, which was lost in mutants with short N-terminal deletions. This antibody, in contrast to the others, failed to immunoprecipitate the native 22K protein, indicating that the N terminus of this protein is buried in the native molecule and exposed only under the denaturing conditions of Western blotting. In addition, N-terminal deletions that abolished reactivity with monoclonal antibody 112 also inhibited phosphorylation of the 22K protein previously identified at Ser-58 and Ser-61, suggesting that the N terminus is important in regulating the 22K protein phosphorylation status, most likely as a result of its requirement for protein folding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Viral / immunology
  • Antibodies, Viral / metabolism
  • Blotting, Western
  • Epitope Mapping*
  • Gene Deletion
  • Gene Expression Regulation, Viral
  • Humans
  • Mutation
  • Protein Folding*
  • Respiratory Syncytial Virus, Human / chemistry
  • Respiratory Syncytial Virus, Human / genetics
  • Respiratory Syncytial Virus, Human / immunology*
  • Respiratory Syncytial Virus, Human / metabolism
  • Transcription, Genetic
  • Viral Proteins / chemistry*
  • Viral Proteins / genetics
  • Viral Proteins / immunology
  • Viral Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Viral Proteins