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TitleStructural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.
Journal, issue, pagesMol Cell, Vol. 80, Issue 1, Page 72-86.e7, Year 2020
Publish dateOct 1, 2020
AuthorsMelanie A McDowell / Michael Heimes / Francesco Fiorentino / Shahid Mehmood / Ákos Farkas / Javier Coy-Vergara / Di Wu / Jani Reddy Bolla / Volker Schmid / Roger Heinze / Klemens Wild / Dirk Flemming / Stefan Pfeffer / Blanche Schwappach / Carol V Robinson / Irmgard Sinning /
PubMed AbstractMembrane protein biogenesis faces the challenge of chaperoning hydrophobic transmembrane helices for faithful membrane insertion. The guided entry of tail-anchored proteins (GET) pathway targets and ...Membrane protein biogenesis faces the challenge of chaperoning hydrophobic transmembrane helices for faithful membrane insertion. The guided entry of tail-anchored proteins (GET) pathway targets and inserts tail-anchored (TA) proteins into the endoplasmic reticulum (ER) membrane with an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively). Here, we present cryo-electron microscopy reconstructions, native mass spectrometry, and structure-based mutagenesis of human WRB/CAML/TRC40 and yeast Get1/Get2/Get3 complexes. Get3 binding to the membrane insertase supports heterotetramer formation, and phosphatidylinositol binding at the heterotetramer interface stabilizes the insertase for efficient TA insertion in vivo. We identify a Get2/CAML cytoplasmic helix that forms a "gating" interaction with Get3/TRC40 important for TA insertion. Structural homology with YidC and the ER membrane protein complex (EMC) implicates an evolutionarily conserved insertion mechanism for divergent substrates utilizing a hydrophilic groove. Thus, we provide a detailed structural and mechanistic framework to understand TA membrane insertion.
External linksMol Cell / PubMed:32910895
MethodsEM (single particle)
Resolution4.2 - 14.0 Å
Structure data

EMDB-10266, PDB-6so5:
Homo sapiens WRB/CAML heterotetramer in complex with a TRC40 dimer
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-11607:
Saccharomyces cerevisiae Get1/Get2 heterotetramer in complex with a Get3 dimer
Method: EM (single particle) / Resolution: 14.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • homo sapiens (human)
  • Saccharomyces cerevisiae (brewer's yeast)
KeywordsMEMBRANE PROTEIN / ER / insertase / complex / tail-anchor

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