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TitleDesigned miniproteins potently inhibit and protect against MERS-CoV.
Journal, issue, pagesCell Rep, Vol. 44, Issue 6, Page 115760, Year 2025
Publish dateJun 24, 2025
AuthorsRobert J Ragotte / M Alejandra Tortorici / Nicholas J Catanzaro / Amin Addetia / Brian Coventry / Heather M Froggatt / Jimin Lee / Cameron Stewart / Jack T Brown / Inna Goreshnik / Jeremiah N Sims / Lukas F Milles / Basile I M Wicky / Matthias Glögl / Stacey Gerben / Alex Kang / Asim K Bera / William Sharkey / Alexandra Schäfer / Jack R Harkema / Ralph S Baric / David Baker / David Veesler /
PubMed AbstractMiddle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic pathogen with a 36% case-fatality rate in humans. No vaccines or specific therapeutics are currently approved for use in humans ...Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic pathogen with a 36% case-fatality rate in humans. No vaccines or specific therapeutics are currently approved for use in humans or the camel host reservoir. Here, we computationally designed monomeric and homo-oligomeric miniproteins that bind with high affinity to the MERS-CoV spike (S) glycoprotein, the main target of neutralizing antibodies and vaccine development. We show that these miniproteins broadly neutralize a panel of MERS-CoV S variants, spanning the known antigenic diversity of this pathogen, by targeting a conserved site in the receptor-binding domain (RBD). The miniproteins directly compete with binding of the dipeptidylpeptidase 4 (DPP4) receptor to MERS-CoV S, thereby blocking viral attachment to the host entry receptor and subsequent membrane fusion. Intranasal administration of a lead miniprotein provides prophylactic protection against stringent MERS-CoV challenge in mice, motivating its future clinical development as a next-generation countermeasure against this virus with pandemic potential.
External linksCell Rep / PubMed:40450691 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.85 - 3.3 Å
Structure data

EMDB-46947: Designed miniproteins potently inhibit and protect against MERS_CoV (Global refinement of MERS_CoV_S RBD in complex with miniprotein cb3_GSG_SB175, linker 1)
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-46952: Designed miniproteins potently inhibit and protect against MERS_CoV. MERS_CoV S in complex with cb3_GSG_SB175, linker 1. Global refinement, three RBDs engaged.
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-46955: Designed miniproteins potently inhibit and protect against MERS_CoV. MERS_CoV S in complex with cb3_GGGSGGGS_SB175, linker 7. Global refinement.
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-46957: Designed miniproteins potently inhibit and protect against MERS_CoV. MERS-CoV S in complex with cb3_GGGSGGGS_SB175B175, linker 7(Global refinement after focused classification)
Method: EM (single particle) / Resolution: 3.0 Å

46960

9dkk

EMDB-46960, PDB-9dkk:
Designed miniproteins potently inhibit and protect against MERS-CoV. MERS-CoV S in complex with miniprotein cb3_GGGSGGGS_SB175, linker 7 (Local refinement of two RBDs and 2 miniproteins)
Method: EM (single particle) / Resolution: 3.3 Å

PDB-9c7z:
Hallucinated C3 protein assembly HALC3_919
Method: X-RAY DIFFRACTION / Resolution: 2.1 Å

PDB-9dgo:
Designed miniproteins potently inhibit and protect against MERS-CoV. Crystal structure of MERS-CoV S RBD in complex with miniprotein cb3
Method: X-RAY DIFFRACTION / Resolution: 1.85 Å

Chemicals

ChemComp-PG4:
TETRAETHYLENE GLYCOL / precipitant*YM

ChemComp-HOH:
WATER

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

Source
  • middle east respiratory syndrome-related coronavirus
  • Homo sapiens (human)
  • synthetic construct (others)
KeywordsDE NOVO PROTEIN / trimer / hallucination / de novo / VIRAL PROTEIN / MERS-CoV / miniproteins / neutralization / fusion / cryo-EM / biolayer interferometry / MERS-CoV strains / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease / SSGCID / Coronavirus / betacoronavirus / single particle cryo-EM / neutralization assays / binding assays / miniproteins inhibitors / fusion assays / in vivo protection

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