6QOZ

CryoEM reconstruction of Cowpea Mosaic Virus (CPMV) bound to an Affimer reagent

Summary for 6QOZ

• Entry DOI: 10.2210/pdb6qoz/pdb
• EMDB information: 4610
• Descriptor: RNA2 polyprotein, Cowpea mosaic virus large subunit, Affimer binding protein (3 entities in total)
• Functional Keywords: virus, cpmv, comoviridae, picornavirales, affimer reagent, antibody-like
• Biological source: Cowpea mosaic virus (CPMV); More
• Total number of polymer chains: 9
• Total formula weight: 256566.67

Authors

Hesketh, E.L.,Tiede, C.,Adamson, H.,Adams, T.L.,Byrne, M.J.,Meshcheriakova, Y.,Lomonossoff, G.P.,Kruse, I.,McPherson, M.J.,Tomlinson, D.C.,Ranson, N.A. (deposition date: 2019-02-13, release date: 2019-12-18, Last modification date: 2024-05-15)

Primary citation

Hesketh, E.L.,Tiede, C.,Adamson, H.,Adams, T.L.,Byrne, M.J.,Meshcheriakova, Y.,Kruse, I.,McPherson, M.J.,Lomonossoff, G.P.,Tomlinson, D.C.,Ranson, N.A.
Affimer reagents as tools in diagnosing plant virus diseases.
Sci Rep, 9:7524-7524, 2019

PubMed Abstract: Plant viruses can cause devastating losses to agriculture and are therefore a major threat to food security. The rapid identification of virally-infected crops allowing containment is essential to limit such threats, but plant viral diseases can be extremely challenging to diagnose. An ideal method for plant virus diagnosis would be a device which can be implemented easily in the field. Such devices require a binding reagent that is specific for the virus of interest. We chose to investigate the use of Affimer reagents, artificial binding proteins and a model plant virus Cowpea Mosaic virus (CPMV) empty virus like particles (eVLPs). CPMV-eVLP mimic the morphology of wild-type (WT) CPMV but lack any infectious genomic material and so do not have biocontainment issues. We have produced and purified an Affimer reagent selected for its ability to bind to CPMV-eVLP and have shown that the selected Affimer also specifically binds to WT CPMV. We have produced a 3.4 Å structure of WT CPMV bound to the Affimer using cryo-electron microscopy. Finally, we have shown that this Affimer is capable of reliably detecting the virus in crude extracts of CPMV-infected leaves and can therefore form the basis for the future development of diagnostic tests.

PubMed: 31101847
DOI: 10.1038/s41598-019-43945-6

Experimental method

ELECTRON MICROSCOPY (3.4 Å)