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TitleCryo-electron microscopy reveals a single domain antibody with a unique binding epitope on fibroblast activation protein alpha.
Journal, issue, pagesRSC Chem Biol, Vol. 6, Issue 5, Page 780-787, Year 2025
Publish dateMay 8, 2025
AuthorsZhen Xu / Akesh Sinha / Darpan N Pandya / Nicholas J Schnicker / Thaddeus J Wadas /
PubMed AbstractFibroblast activation protein alpha (FAP) is a serine protease that is expressed at basal levels in benign tissues but is overexpressed in a variety of pathologies, including cancer. Despite this ...Fibroblast activation protein alpha (FAP) is a serine protease that is expressed at basal levels in benign tissues but is overexpressed in a variety of pathologies, including cancer. Despite this unique expression profile, designing functional diagnostic and therapeutic agents that effectively target this biomarker remains elusive. Here we report the structural characterization of the interaction between a novel single domain antibody (sdAb), I3, and FAP using cryo-electron microscopy. The reconstructions were determined to a resolution of 2.7 Å and contained two distinct populations; one I3 bound and two I3 molecules bound to the FAP dimer. In both cases, the sdAb bound a unique epitope that was distinct from the active site of the enzyme. Furthermore, this report describes the rational mutation of specific residues within the complementarity determining region 3 (CDR3) loop to enhance affinity and selectivity of the I3 molecule for FAP. This report represents the first sdAb-FAP structure to be described in the literature.
External linksRSC Chem Biol / PubMed:39975582 / PubMed Central
MethodsEM (single particle)
Resolution2.7 Å
Structure data

47215

9dvq

EMDB-47215, PDB-9dvq:
Cryo-EM structure of Human Fibroblast Activation Protein alpha dimer with one SUMO-I3 VHHs bound
Method: EM (single particle) / Resolution: 2.7 Å

47216

9dvr

EMDB-47216, PDB-9dvr:
Cryo-EM structure of Human Fibroblast Activation Protein alpha dimer with two SUMO-I3 VHHs bound
Method: EM (single particle) / Resolution: 2.7 Å

Chemicals

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

Source
  • homo sapiens (human)
  • lama glama (llama)
KeywordsHYDROLASE / Fibroblast activation protein / single domain antibody / Protease / VHH

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