A Fungal Defensin Targets the SARS−CoV−2 Spike Receptor−Binding Domain

Gao, Bin; Zhu, Shunyi

doi:10.3390/jof7070553

Cited by 15 publications

(20 citation statements)

References 52 publications

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“… 34 Earlier studies also demonstrated the binding of a fungal defensin into the spike RBD from SARS‐CoV‐2 . 35 However, unlike our study, in both these cases, the defensin alpha‐helix was not engineered to mimic the ACE2 alpha helix1, and perhaps for that reason, their binding affinities for SARS‐CoV‐2 spike protein were observed at micromolar ranges.…”

Section: Discussionmentioning

confidence: 70%

“…Recently, a human beta‐defensin expressed in the human respiratory epithelium was shown to bind to the spike RDB and inhibit SARS‐CoV‐2 pseudovirus infection on ACE2 expressing HEK 293Tcells 34 . Earlier studies also demonstrated the binding of a fungal defensin into the spike RBD from SARS‐CoV‐2 35 . However, unlike our study, in both these cases, the defensin alpha‐helix was not engineered to mimic the ACE2 alpha helix1, and perhaps for that reason, their binding affinities for SARS‐CoV‐2 spike protein were observed at micromolar ranges.…”

Section: Discussionmentioning

confidence: 99%

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Engineering defensin α‐helix to produce high‐affinitySARS‐CoV‐2 spike protein binding ligands

et al. 2022

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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 mimetic 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 (2-5 kDa) and highly stable proteins containing solvent-exposed alphahelix, conformationally constrained by two disulfide bonds. Therefore, we engineered the amino acid residues on the constrained alpha-helix of defensins to mimic the critical residues on the ACE2 helix 1 that interact with the SARS-CoV-2 spike protein. The engineered proteins (h-deface2, p-deface2, and p-deface2-MUT) were soluble and purified to homogeneity with a high yield from a bacterial expression system. The proteins demonstrated exceptional thermostability (Tm 70.7 C), high-affinity binding to the spike protein with apparent K d values of 54.4 ± 11.3, 33.5 ± 8.2, and 14.4 ± 3.5 nM for h-deface2, p-deface2, and p-deface2-MUT, respectively, and were used in a diagnostic assay that detected SARS-CoV-2 neutralizing antibodies. This work addresses the challenge of developing helical ACE2 mimetics by demonstrating that defensins provide promising scaffolds to engineer alpha-helices in a constrained form for designing of high-affinity ligands.

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Section: Discussionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Engineering defensin α‐helix to produce high‐affinitySARS‐CoV‐2 spike protein binding ligands

et al. 2022

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“…Recently, a human beta-defensin expressed in the human respiratory epithelium was shown to bind to the Spike RDB and inhibit SARS-CoV-2 pseudovirus infection on ACE2 expressing HEK 293Tcells (33). Earlier studies also demonstrated the binding of a fungal defensin into the Spike RBD from SARS-CoV-2 (34). However, unlike our study, in both these cases, the defensin alpha-helix was not engineered to mimic the ACE2 alpha helix1, and perhaps for that reason, their binding affinities for SARS-CoV-2 Spike protein were observed at micromolar ranges.…”

Section: Discussionmentioning

confidence: 99%

Engineering Defensin α-helix to produce high-affinity SARS-CoV-2 Spike protein binding ligands

Fernandes

Gomes

Magalhães

et al. 2022

Preprint

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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.

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“…For this reason, mutants are designed and developed by genetic engineering, which are then tested for better performance as medicine in treatment of various diseases [152]. An example is the fungal defensin Micasin from Microsporum canis, where a synthetically synthesized truncated version of the peptide binds the receptor-binding domain (RBD) of the COVID-19 spike protein six times better than the original mature peptide [153], potentially preventing or limiting the virus by penetrating into the cells.…”

Section: Production Of Anti-microbial Peptidesmentioning

confidence: 99%

Fungal Cell Factories for Efficient and Sustainable Production of Proteins and Peptides

Lübeck

2022

Microorganisms

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Filamentous fungi are a large and diverse taxonomically group of microorganisms found in all habitats worldwide. They grow as a network of cells called hyphae. Since filamentous fungi live in very diverse habitats, they produce different enzymes to degrade material for their living, for example hydrolytic enzymes to degrade various kinds of biomasses. Moreover, they produce defense proteins (antimicrobial peptides) and proteins for attaching surfaces (hydrophobins). Many of them are easy to cultivate in different known setups (submerged fermentation and solid-state fermentation) and their secretion of proteins and enzymes are often much larger than what is seen from yeast and bacteria. Therefore, filamentous fungi are in many industries the preferred production hosts of different proteins and enzymes. Edible fungi have traditionally been used as food, such as mushrooms or in fermented foods. New trends are to use edible fungi to produce myco-protein enriched foods. This review gives an overview of the different kinds of proteins, enzymes, and peptides produced by the most well-known fungi used as cell factories for different purposes and applications. Moreover, we describe some of the challenges that are important to consider when filamentous fungi are optimized as efficient cell factories.

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A Fungal Defensin Targets the SARS−CoV−2 Spike Receptor−Binding Domain

Cited by 15 publications

References 52 publications

Engineering defensin α‐helix to produce high‐affinitySARS‐CoV‐2 spike protein binding ligands

Engineering defensin α‐helix to produce high‐affinitySARS‐CoV‐2 spike protein binding ligands

Engineering Defensin α-helix to produce high-affinity SARS-CoV-2 Spike protein binding ligands

Fungal Cell Factories for Efficient and Sustainable Production of Proteins and Peptides

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