RT Journal Article
SR Electronic
T1 Engineering Defensin α-helix to produce high-affinity SARS-CoV-2 Spike protein binding ligands
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.02.09.479781
DO 10.1101/2022.02.09.479781
A1 Leonardo Antônio Fernandes
A1 Anderson Albino Gomes
A1 Maria de Lourdes Borba Magalhães
A1 Partha Ray
A1 Gustavo Felippe da Silva
YR 2022
UL http://biorxiv.org/content/early/2022/02/10/2022.02.09.479781.abstract
AB The binding of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Spike protein to the Angiotensin-Converting Enzyme 2 (ACE2) receptor expressed on the host cells is a critical initial step for viral infection. This interaction is blocked through competitive inhibition by soluble ACE2 protein. Therefore, developing high-affinity and cost-effective ACE2 peptidomimetic ligands that disrupt this protein-protein interaction is a promising strategy for viral diagnostics and therapy. We employed human and plant defensins, a class of small and highly stable proteins, and engineered the amino acid residues on its conformationally constrained alpha-helices to mimic the critical residues on the ACE2 helix 1 that interacts with the Spike-protein. The engineered proteins were soluble and purified to homogeneity with high yield from a bacterial expression system. The proteins demonstrated exceptional thermostability, high-affinity binding to the Spike protein with dissociation constants in the low nanomolar range, and were used in a diagnostic assay that detected SARS-CoV-2 neutralizing antibodies. This work addresses the challenge of developing helical peptidomimetics by demonstrating that defensins provide promising scaffolds to engineer alpha-helices in a constrained form for designing high-affinity ligands.Broad audience statement The engineered proteins developed in this study are cost-effective and highly stable reagents for SARS-CoV-2 detection. These features may allow large-scale and cost-effective production of diagnostic tests to assist COVID-19 diagnostic and prevention.Competing Interest StatementThe authors intend to file a provisional patent application regarding the engineered proteins.(COVID-19)Coronavirus Disease 2019(SARS-CoV-2)severe acute respiratory syndrome coronavirus 2(RBD)receptor binding domain(ECD)extracellular domain(ACE2)Angiotensin Converting Enzyme 2