Single-molecule conformational dynamics of viroporin ion channels regulated by lipid-protein interactions

Highlights

• p7 viroporin forms pH and membrane dependent nanometer-size ion channels.

• AFM also shows much wider pH regulated channels (tens of nanometers).

• CSFV p7-induced pores are ruled by equilibrium conformational dynamics.

• Dynamics of crowded biomembrane systems were seen depending on lipid composition.

• Sub-nanometric channels may be involved in virus propagation.

• Virus pathogenicity may be related to the larger pores (1 ~ 10 nm in diameter).

Abstract

Classic swine fever is a highly contagious and often fatal viral disease that is caused by the classical swine fever virus (CSFV). Protein p7 of CFSV is a prototype of viroporin, a family of small, highly hydrophobic proteins postulated to modulate virus-host interactions during the processes of virus entry, replication and assembly. It has been shown that CSFV p7 displays substantial ion channel activity when incorporated into membrane systems, but a deep rationalization of the size and dynamics of the induced pores is yet to emerge. Here, we use high-resolution conductance measurements and current fluctuation analysis to demonstrate that CSFV p7 channels are ruled by equilibrium conformational dynamics involving protein-lipid interactions. Atomic force microscopy (AFM) confirms the existence of a variety of pore sizes and their tight regulation by solution pH. We conclude that p7 viroporin forms subnanometric channels involved in virus propagation, but also much larger pores (1–10 nm in diameter) with potentially significant roles in virus pathogenicity. Our findings provide new insights into the sources of noise in protein electrochemistry and demonstrate the existence of slow complex dynamics characteristic of crowded systems like biomembrane surfaces.