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TitleImplications for tetraspanin-enriched microdomain assembly based on structures of CD9 with EWI-F.
Journal, issue, pagesLife Sci Alliance, Vol. 3, Issue 11, Year 2020
Publish dateSep 21, 2020
AuthorsWout Oosterheert / Katerina T Xenaki / Viviana Neviani / Wouter Pos / Sofia Doulkeridou / Jip Manshande / Nicholas M Pearce / Loes Mj Kroon-Batenburg / Martin Lutz / Paul Mp van Bergen En Henegouwen / Piet Gros /
PubMed AbstractTetraspanins are eukaryotic membrane proteins that contribute to a variety of signaling processes by organizing partner-receptor molecules in the plasma membrane. How tetraspanins bind and cluster ...Tetraspanins are eukaryotic membrane proteins that contribute to a variety of signaling processes by organizing partner-receptor molecules in the plasma membrane. How tetraspanins bind and cluster partner receptors into tetraspanin-enriched microdomains is unknown. Here, we present crystal structures of the large extracellular loop of CD9 bound to nanobodies 4C8 and 4E8 and, the cryo-EM structure of 4C8-bound CD9 in complex with its partner EWI-F. CD9-EWI-F displays a tetrameric arrangement with two central EWI-F molecules, dimerized through their ectodomains, and two CD9 molecules, one bound to each EWI-F transmembrane helix through CD9-helices h3 and h4. In the crystal structures, nanobodies 4C8 and 4E8 bind CD9 at loops C and D, which is in agreement with the 4C8 conformation in the CD9-EWI-F complex. The complex varies from nearly twofold symmetric (with the two CD9 copies nearly anti-parallel) to ca. 50° bent arrangements. This flexible arrangement of CD9-EWI-F with potential CD9 homo-dimerization at either end provides a "concatenation model" for forming short linear or circular assemblies, which may explain the occurrence of tetraspanin-enriched microdomains.
External linksLife Sci Alliance / PubMed:32958604 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.33 - 8.6 Å
Structure data

EMDB-11053:
Cryo-EM density map of nanobody-4C8 bound, human CD9 in complex with EWI-F-deltaIg1-5
Method: EM (single particle) / Resolution: 8.6 Å

PDB-6rlr:
Crystal structure of CD9 large extracellular loop
Method: X-RAY DIFFRACTION / Resolution: 2.0 Å

PDB-6z1v:
Structure of the EC2 domain of CD9 in complex with nanobody 4E8
Method: X-RAY DIFFRACTION / Resolution: 1.33 Å

PDB-6z1z:
Structure of the anti-CD9 nanobody 4C8
Method: X-RAY DIFFRACTION / Resolution: 1.7 Å

PDB-6z20:
Structure of the EC2 domain of CD9 in complex with nanobody 4C8
Method: X-RAY DIFFRACTION / Resolution: 2.68 Å

Chemicals

ChemComp-HOH:
WATER

ChemComp-PGE:
TRIETHYLENE GLYCOL

ChemComp-EDO:
1,2-ETHANEDIOL

ChemComp-ACY:
ACETIC ACID

ChemComp-GOL:
GLYCEROL

ChemComp-CL:
Unknown entry

Source
  • homo sapiens (human)
  • lama glama (llama)
KeywordsIMMUNE SYSTEM / CD9 antigen / tetraspanin / CELL ADHESION / Antibody-antigen complex / EC2 domain / nanobody / CD9-binding / CD9

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