Evidence for a polytopic form of the E1 envelope glycoprotein of Hepatitis C virus

Christopher T. Migliaccio, Kathryn E. Follis, Yoshiharu Matsuura, Jack H. Nunberg

Research output: Contribution to journalArticlepeer-review

Abstract

The polyprotein precursor of the Hepatitis C virus (HCV) contains multiple membrane-spanning domains that define the membrane topology and subsequent maturation of the viral structural proteins. In order to examine the biogenesis of the E1-E2 heterodimeric complex, we inserted an affinity tag (S-peptide) at specific locations within the envelope glycoproteins. In particular, and based on the prediction that the E1 glycoprotein may be able to assume a polytopic topology containing two membrane-spanning domains, we inserted the affinity tag within a putative cytoplasmic loop of the E1 glycoprotein. The HCV structural polyprotein containing this tag (at amino acids 295/296) was highly expressed and able to form a properly processed and noncovalently associated E1-E2 complex. This complex was bound by murine and conformation-dependent human monoclonal antibodies (MAbs) comparably to the native untagged complex. In addition, MAb recognition was retained upon reconstituting the tagged E1-E2 complex in lipid membrane as topologically constrained proteoliposomes. Our findings are consistent with the model of a topologically flexible E1 glycoprotein that is able to adopt a polytopic form. This form of the E1-E2 complex may be important in the HCV life cycle and in pathogenesis.

Original languageEnglish
Pages (from-to)47-57
Number of pages11
JournalVirus Research
Volume105
Issue number1
DOIs
StatePublished - Sep 15 2004

Keywords

  • Affinity tag
  • E1-E2 complex
  • Envelope glycoprotein
  • Hepatitis C virus
  • Membrane topology
  • Proteoliposome

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