Quasi-crystalline Arrangement of Human Rhinovirus 2 on Model Cell Membranes

Abstract

We used dynamic force microscopy to study the attachment of human rhinovirus 2 (HRV2) particles to model membranes. A supported lipid bilayer containing Ni 2+nitrilotriacetate (NTA)-lipids as functional groups in the outer leaflet at high surface density was assembled on mica and showed a flat extended structure of 3 nm in height. The recombinant very-low density lipoprotein receptor fragment MBP-VLDLR1-8, consisting of the ligand binding domain (all eight complement type A repeats) fused at the N-terminus to maltose binding protein (MBP) and containing a hexahistidine (His6)-tag at its C-terminus, was attached to the membranes via the His6-Ni 2+ NTA bond and revealed monolayer density. The protein-lipid layer was then used as a model to investigate virus binding to its receptor in a planar membrane. The viral particles attached in a tightly packed, long range order quasi-crystalline arrangement. This assembly is thus ideally suited for studies of viral dynamics and recognition under physiological conditions.

Dynamic force microscopy for imaging of viruses under physiological conditions

“…In a previous study, HRV2 bound to an artificial cell membrane containing derivatives of the very-low density lipoprotein receptor was studied using MACmode AFM (24). The presence of the receptor resulted in specific, tight immobilization of HRV2.…”

“…In a previous study we used DFM to investigate the binding and the arrangement of the minor-group virus HRV2 on artificial cell membranes, mimicking an early step of virus entry into living cells (24). In our recent study, DFM was used to image RNA directly in the process of extrusion from HRV2 (25).…”