Development and structure-guided characterization of a novel ACE2-binding macrocyclic peptide
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Abstract
Abstract Angiotensin-converting enzyme 2 (ACE2) is a key node in the protective axis of the renin-angiotensin-aldosterone system (RAAS) for blood pressure and hydroelectrolyte regulation, and the main protein receptor recognized by the spike glycoproteins of the severe acute respiratory syndrome (SARS) coronaviruses (CoV) SARS-CoV and SARS-CoV-2. We identified the macrocyclic peptide WJL-63, developed using mRNA display, with high ACE2-binding affinity. The peptide was characterized in vitro in terms of purity, stability, hydrophilicity and ACE2 binding. The crystal structure of the extracellular region of ACE2 in complex with the peptide at 2.2 Å resolution was elucidated. The structure revealed a binding mode in which WJL-63 is accommodated towards one side of the wide catalytic cleft of the ACE2 peptidase domain, with no direct contact to the conserved zinc ion site. WJL-63 residues Q4, R7, R11 and R14 anchor the peptide deep inside the binding pocket. The opposite edges of the peptide were found to be in contact with subdomain 1 and subdomain 2 of the peptidase domain. This upright binding mode requires an open ACE2 conformation, in contrast to small molecule carboxypeptidase inhibitors, which typically bind to the closed conformation of the enzyme. As a consequence of the open conformation binding mode, the front edge of WJL-63 is accessible for modification such as the herein reported conjugation of a chelator for radiometal labeling. The radiolabeled DOTA-WJL-63 was evaluated on ACE2-transfected HEK cells on which it revealed relatively strong binding with a K D value of 90 ± 28 nM. WJL-63 provides a strong basis for the development of new classes of compounds for the modulation of ACE2 conformation, and for the development of imaging agents for the visualization of ACE2, for example in fluorescence or electron microscopy, or positron emission tomography (PET) imaging.
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- Authors
- Benoit, R. M., Wang, J., Beyer, D., Abbas, A., Rodrigues, M. J., Wieser, M. M., Deupi, X., Müller, C., Suga, H., & Bode, J. W.
- Affiliations
- Paul Scherrer Institute (CH)
- ETH Zurich (CH)
- The University of Tokyo (JP)
- Diamond Light Source (GB)
- Corresponding author
- Benoit, Roger M.
- ORCID
- Venue
- Journal of Structural Biology: X
- Publisher
- Elsevier BV
- Published
- 2026-06
- Volume
- 13
- Pages
- 100145
- ISSN
- 2590-1524 (ISSN-L)
- OpenAlex
- W4417116900
- Cited by
- 0
- References
- 37
- FWCI
- 0.00
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Journal of Structural Biology X (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by)
- Repository for Publications and Research Data (ETH Zurich) (publishedVersion, cc-by)
- DORA PSI (Paul Scherrer Institute) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- ETH Zürich Research Collection (cc-by)
- Funders
- Promedica Foundation
- Swiss National Science Foundation (CRSK-3_190414, 310030_188978)
- Paul Scherrer Institut
- Japan Society for the Promotion of Science (JP20H05618)
- Topics
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Renin-Angiotensin System Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Health Sciences › Medicine › Oncology › Peptidase Inhibition and Analysis
- Keywords
- ACE2, macrocyclic peptides, mRNA display, peptide-peptide binding, crystal structure, SARS-CoV-2, SARS-CoV, renin-angiotensin-aldosterone system, radiolabeling, DOTA conjugates, PET imaging agents
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