Correlation Engine 2.0
Clear Search sequence regions

Sizes of these terms reflect their relevance to your search.

The HIV capsid is a multifunctional protein capsule that mediates the delivery of the viral genetic material into the nucleus of the target cell. Host cell proteins bind to a number of repeating binding sites on the capsid to regulate steps in the replication cycle. Here, we develop a fluorescence fluctuation spectroscopy method using self-assembled capsid particles as the bait to screen for fluorescence-labeled capsid-binding analytes ("prey" molecules) in solution. The assay capitalizes on the property of the HIV capsid as a multivalent interaction platform, facilitating high sensitivity detection of multiple prey molecules that have accumulated onto capsids as spikes in fluorescence intensity traces. By using a scanning stage, we reduced the measurement time to 10 s without compromising on sensitivity, providing a rapid binding assay for screening libraries of potential capsid interactors. The assay can also identify interfaces for host molecule binding by using capsids with defects in known interaction interfaces. Two-color coincidence detection using the fluorescent capsid as the bait further allows the quantification of binding levels and determination of binding affinities. Overall, the assay provides new tools for the discovery and characterization of molecules used by the HIV capsid to orchestrate infection. The measurement principle can be extended for the development of sensitive interaction assays, utilizing natural or synthetic multivalent scaffolds as analyte-binding platforms.


Derrick Lau, James C Walsh, Claire F Dickson, Andrew Tuckwell, Jeffrey H Stear, Dominic J B Hunter, Akshay Bhumkar, Vaibhav Shah, Stuart G Turville, Emma Sierecki, Yann Gambin, Till Böcking, David A Jacques. Rapid HIV-1 Capsid Interaction Screening Using Fluorescence Fluctuation Spectroscopy. Analytical chemistry. 2021 Mar 02;93(8):3786-3793

Expand section icon Mesh Tags

Expand section icon Substances

PMID: 33593049

View Full Text