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Structure paper

TitleStructural insights into Frizzled3 through nanobody modulators.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 7228, Year 2024
Publish dateAug 22, 2024
AuthorsJames Hillier / Yuguang Zhao / Loic Carrique / Tomas Malinauskas / Reinis R Ruza / Tao-Hsin Chang / Gangshun Yi / Helen M E Duyvesteyn / Jing Yu / Weixian Lu / Els Pardon / Jan Steyaert / Yanan Zhu / Tao Ni / E Yvonne Jones /
PubMed AbstractThe Wnt receptor Frizzled3 (FZD3) is important for brain axonal development and cancer progression. We report structures of FZD3 in complex with extracellular and intracellular binding nanobodies (Nb) ...The Wnt receptor Frizzled3 (FZD3) is important for brain axonal development and cancer progression. We report structures of FZD3 in complex with extracellular and intracellular binding nanobodies (Nb). The crystal structure of Nb8 in complex with the FZD3 cysteine-rich domain (CRD) reveals that the nanobody binds at the base of the lipid-binding groove and can compete with Wnt5a. Nb8 fused with the Dickkopf-1 C-terminal domain behaves as a FZD3-specific Wnt surrogate, activating β-catenin signalling. The cryo-EM structure of FZD3 in complex with Nb9 reveals partially resolved density for the CRD, which exhibits positional flexibility, and a transmembrane conformation that resembles active GPCRs. Nb9 binds to the cytoplasmic region of FZD3 at the putative Dishevelled (DVL) or G protein-binding site, competes with DVL binding, and inhibits GαS coupling. In combination, our FZD3 structures with nanobody modulators map extracellular and intracellular interaction surfaces of functional, and potentially therapeutic, relevance.
External linksNat Commun / PubMed:39174501 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.76 - 2.9 Å
Structure data

18680

8qw4

EMDB-18680, PDB-8qw4:
FZD3 in complex with nanobody 9
Method: EM (single particle) / Resolution: 2.9 Å

PDB-8q7o:
Crystal structure of the FZD3 cysteine-rich domain in complex with a nanobody (14478)
Method: X-RAY DIFFRACTION / Resolution: 1.76 Å

Chemicals

ChemComp-DMS:
DIMETHYL SULFOXIDE / DMSO, precipitant*YM

ChemComp-GOL:
GLYCEROL

ChemComp-CIT:
CITRIC ACID

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

ChemComp-SO4:
SULFATE ION

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
  • lama glama (llama)
KeywordsSIGNALING PROTEIN / Frizzled / WNT / GPCR / Nanobody / Complex / CELL ADHESION / FZD3 Nanobody

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