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TitleCryo-EM study of patched in lipid nanodisc suggests a structural basis for its clustering in caveolae.
Journal, issue, pagesStructure, Vol. 29, Issue 11, Page 1286-11294.e6, Year 2021
Publish dateNov 4, 2021
AuthorsYitian Luo / Guoyue Wan / Xiang Zhang / Xuan Zhou / Qiuwen Wang / Jialin Fan / Hongmin Cai / Liya Ma / Hailong Wu / Qianhui Qu / Yao Cong / Yun Zhao / Dianfan Li /
PubMed AbstractThe 12-transmembrane protein Patched (Ptc1) acts as a suppressor for Hedgehog (Hh) signaling by depleting sterols in the cytoplasmic membrane leaflet that are required for the activation of ...The 12-transmembrane protein Patched (Ptc1) acts as a suppressor for Hedgehog (Hh) signaling by depleting sterols in the cytoplasmic membrane leaflet that are required for the activation of downstream regulators. The positive modulator Hh inhibits Ptc1's transporter function by binding to Ptc1 and its co-receptors, which are locally concentrated in invaginated microdomains known as caveolae. Here, we reconstitute the mouse Ptc1 into lipid nanodiscs and determine its structure using single-particle cryoelectron microscopy. The structure is overall similar to those in amphipol and detergents but displays various conformational differences in the transmembrane region. Although most particles show monomers, we observe Ptc1 dimers with distinct interaction patterns and different membrane curvatures, some of which are reminiscent of caveolae. We find that an extramembranous "hand-shake" region rich in hydrophobic and aromatic residues mediates inter-Ptc1 interactions under different membrane curvatures. Our data provide a plausible framework for Ptc1 clustering in the highly curved caveolae.
External linksStructure / PubMed:34174188
MethodsEM (single particle)
Resolution3.5 - 3.64 Å
Structure data

31753

7v6y

EMDB-31753, PDB-7v6y:
Cryo-EM structure of Patched in lipid nanodisc - the wildtype, 3.5 angstrom (re-processed with dataset of 7dzq)
Method: EM (single particle) / Resolution: 3.5 Å

31754

7v6z

EMDB-31754, PDB-7v6z:
Cryo-EM structure of Patched1 (V1084A mutant) in lipid nanodisc, 3.64 angstrom (reprocessed with the dataset of 7dzp)
Method: EM (single particle) / Resolution: 3.64 Å

Chemicals

ChemComp-CLR:
CHOLESTEROL

ChemComp-5VI:
(2S)-2-azanyl-3-[[(2S)-3-butanoyloxy-2-dec-9-enoyloxy-propoxy]-oxidanyl-phosphoryl]oxy-propanoic acid

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

Source
  • mus musculus (house mouse)
KeywordsMEMBRANE PROTEIN / Caveolae / Hedgehog signaling / Lipid nanodisc / Patched / Ptc1 dimer / Thermostable mutant.

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