Computational and Experimental Identification of Potential Neutralizing Peptides Derived from Human ACE2 Against...

Area of Science:

• Virology
• Immunology
• Computational Biology

Background:

• The human angiotensin-converting enzyme 2 (hACE2) serves as the primary receptor for SARS-CoV-2 entry.
• ACE2 polymorphisms can influence viral affinity, but their impact on infection is not fully understood.
• Previous work identified key residues in SARS-CoV-2 Spike protein and hACE2 critical for binding.

Purpose of the Study:

• To explore hACE2-derived neutralizing peptides as a therapeutic strategy against SARS-CoV-2.
• To investigate the effect of ACE2 polymorphisms on the binding affinity with the SARS-CoV-2 Spike protein receptor-binding domain (RBD).

Main Methods:

• Computational saturation mutagenesis was used to screen all possible ACE2 missense mutations within the ACE2-Wuhan-S1 complex.
• Six vital ACE2 regions for ACE2-RBD interaction were identified computationally.
• Peptides corresponding to these regions were synthesized and tested using pseudotyped viral particles and dot blot assays.
• ACE2-Wuhan-S1 and ACE2-Omicron-S1 complexes were aligned and analyzed for binding patterns.

Main Results:

• Mutations at ACE2 residues D355 and Y41 were predicted to weaken binding affinity, while N330 and D30 enhanced it.
• Six ACE2 regions (19-49, 65-102, 320-333, 348-359, 378-395, 552-563) were identified as vital for ACE2-RBD interaction.
• Three synthesized peptides bound the S protein, and four exhibited inhibitory effects on the Wuhan strain.
• Analysis of Wuhan and Omicron complexes showed similar binding patterns, suggesting peptide efficacy against Omicron variants.

Conclusions:

• hACE2-derived peptides show potential as a therapeutic strategy against SARS-CoV-2, including Omicron variants.
• The study highlights the role of ACE2 polymorphisms in viral entry.
• Strong consistency was observed between computational predictions and experimental outcomes.