Quantitative investigation of the direct interaction between Hemagglutinin and fusion proteins of Peste des petits ruminant virus using surface Plasmon resonance

Virol J. 2018 Jan 22;15(1):21. doi: 10.1186/s12985-018-0933-7.

Abstract

Background: The specific and dynamic interaction between the hemagglutinin (H) and fusion (F) proteins of morbilliviruses is a prerequisite for the conformational rearrangements and membrane fusion during infection process. The two heptad repeat regions (HRA and HRB) of F protein are both important for the triggering of F protein.

Methods: In this study, the direct interactions of Peste des petits ruminants virus (PPRV) H with F, HRA and HRB were quantitatively evaluated using biosensor surface plasmon resonance (SPR).

Results: The binding affinities of immobilized pCMV-HA-H (HA-H) interacted with proteins pCMV-HA-F (HA-F) and pCMV-HA-HRB (HA-HRB) (KD = 1.91 × 10- 8 M and 2.60 × 10- 7 M, respectively) reacted an order of magnitude more strongly than that of pCMV-HA-HRA (HA-HRA) and pCMV-HA-Tp IGFR-LD (HA) (KD = 1.08 × 10- 4 M and 1.43 × 10- 4 M, respectively).

Conclusions: The differences of the binding affinities suggested that HRB is involved in functionally important intermolecular interaction in the fusion process.

Keywords: Expression; Fusion protein; Hemagglutinin; Interactions; Peste des petits ruminanst virus; SPR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetulus
  • Flow Cytometry
  • Gene Expression
  • Hemagglutinins, Viral / chemistry
  • Hemagglutinins, Viral / genetics
  • Hemagglutinins, Viral / metabolism*
  • Kinetics
  • Peste-des-Petits-Ruminants / virology*
  • Peste-des-petits-ruminants virus / physiology*
  • Protein Binding
  • Recombinant Proteins
  • Surface Plasmon Resonance
  • Viral Fusion Proteins / chemistry
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / metabolism*

Substances

  • Hemagglutinins, Viral
  • Recombinant Proteins
  • Viral Fusion Proteins