Exploration of the metal coordination region of concanavalin A for its interaction with human norovirus

Abstract

Rapid methods for the detection and clinical treatment of human norovirus (HuNoV) are needed to control foodborne disease outbreaks, but reliable techniques that are fast and sensitive enough to detect small amounts of HuNoV in food and aquatic environments are not yet available. We explore the interactions between HuNoV and concanavalin A (Con A), which could facilitate the development of a sensitive detection tool for HuNoV. Biophysical studies including hydrogen/deuterium exchange (HDX) mass spectrometry and surface plasmon resonance (SPR) revealed that when the metal coordinated region of Con A, which spans Asp16 to His24, is converted to nine alanine residues (mCon A), the affinity for HuNoV (GII.4) diminishes, demonstrating that this Ca and Mn coordinated region is responsible for the observed virus-protein interaction. The mutated carbohydrate binding region of Con A (mCon A) does not affect binding affinity significantly, indicating that MCR of Con A is a major region of interaction to HuNoV (GII.4). The results further contribute to the development of a HuNoV concentration tool, Con A-immobilized polyacrylate beads (Con A-PAB), for rapid detection of genotypes from genogroups I and II (GI and GII). This method offers many advantages over currently available methods, including a short concentration time. HuNov (GI and GII) can be detected in just 15 min with 90% recovery through Con A-PAB application. In addition, this method can be used over a wide range of pH values (pH 3.0 - 10.0). Overall, this rapid and sensitive detection of HuNoV (GI and GII) will aid in the prevention of virus transmission pathways, and the method developed here may have applicability for other foodborne viral infections.

Chicken GRIFIN: Structural characterization in crystals and in solution

“…In solution, monitoring HDX in peptic fingerprints at 100% sequence coverage, too, confirmed location of the binding site and did not record any significant liganddependent change. Localization of contact site(s) for ligands and monitoring for an impact of the association on solvent accessibility of protein regions are the strengths of this technique [31,76]. Implementing the Asn48Lys change, a hallmark of mammalian GRIFINs, made binding of this mutant protein to tissue sections rather insensitive to lactose presence, intimating this position to be a key site for interactions.…”

A Survey of Analytical Techniques for Noroviruses

“…By SPR, Hurwitz et al [57] identified two scFvs (NJT-R3-A2 and NJT-R3-A3) that could bind to GI.1 and GI.7 VLPs, with Kd values of 27 nM and 49 nM, respectively. Similarly, Kim et al [137] demonstrated that concanavalin A (Con A) can bind with HuNoV (GII.4), with the metal coordination region of Con A identified as a major [129] with permission from the publisher).…”

“…Specifically, the Kd values determined for native Con A and metal coordinating region-mutated Con A by SPR were 71.5 ± 6.7 nM and 22.0 ± 17.0 µM, respectively. By using Con A-immobilized polyacrylate beads, HuNoV (GI and GII) could be rapidly detected in 15 min with 90% recovery over a wide range of pH values (pH 3.0~10.0) [137].…”

“…Specifically, the Kd values determined for native Con A and metal coordinating region-mutated Con A by SPR were 71.5 ± 6.7 nM and 22.0 ± 17.0 μM, respectively. By using Con A-immobilized polyacrylate beads, HuNoV (GI and GII) could be rapidly detected in 15 min with 90% recovery over a wide range of pH values (pH 3.0~10.0) [137]. Furthermore, a localized SPR-based immunofluorescence nanobiosensor developed by Takemura et al [132] was able to achieve a LOD of 0.4 pg/mL of NoV VLPs, by incorporating antibody conjugated AuNPs and quantum dots (QDs).…”

Performance of concanavalin A-immobilized on polyacrylate beads for the detection of human norovirus and hepatitis A virus in fecal specimens

“…However, other foodborne viruses could not recognize HBGA, and not all HuNoV genotypes interact with HBGA (Almand et al, 2017;Singh et al, 2016). Alternatively, concanavalin A (Con A), a carbohydrate-binding lectin, showed a potential for concentrating HuNoV (Kim et al, 2017). Even though the Con A has successfully concentrated HuNoV in artificially contaminated food, it has not been tested for other foodborne viruses on naturally contaminated fecal samples (Kim et al, 2017).…”