Katharina Braunger / Stefan Pfeffer / Shiteshu Shrimal / Reid Gilmore / Otto Berninghausen / Elisabet C Mandon / Thomas Becker / Friedrich Förster / Roland Beckmann /
PubMed Abstract
Protein synthesis, transport, and N-glycosylation are coupled at the mammalian endoplasmic reticulum by complex formation of a ribosome, the Sec61 protein-conducting channel, and ...Protein synthesis, transport, and N-glycosylation are coupled at the mammalian endoplasmic reticulum by complex formation of a ribosome, the Sec61 protein-conducting channel, and oligosaccharyltransferase (OST). Here we used different cryo-electron microscopy approaches to determine structures of native and solubilized ribosome-Sec61-OST complexes. A molecular model for the catalytic OST subunit STT3A (staurosporine and temperature sensitive 3A) revealed how it is integrated into the OST and how STT3-paralog specificity for translocon-associated OST is achieved. The OST subunit DC2 was placed at the interface between Sec61 and STT3A, where it acts as a versatile module for recruitment of STT3A-containing OST to the ribosome-Sec61 complex. This detailed structural view on the molecular architecture of the cotranslational machinery for N-glycosylation provides the basis for a mechanistic understanding of glycoprotein biogenesis at the endoplasmic reticulum.
EMDB-4306: Subtomogram average of unsorted ribosome-translocon complexes from STT3B(-/-) HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4307: Subtomogram average of OST-free ribosome-translocon complexes from STT3B(-/-) HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4308: Subtomogram average of OST-containing ribosome-translocon complexes from STT3B(-/-) HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4309: Subtomogram average of unsorted ribosome-translocon complexes from STT3A(-/-) HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4310: Subtomogram average of OST-free ribosome-translocon complexes from STT3A(-/-) HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4311: Subtomogram average of ribosome-translocon complexes from STT3A(-/-) HEK cells containing an unidentified ER-lumenal component Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4312: Subtomogram average of unsorted ribosome-translocon complexes from wild type HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4313: Subtomogram average of OST-free ribosome-translocon complexes from wild type HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4314: Subtomogram average of OST-containing ribosome-translocon complexes from wild type HEK cells Method: EM (subtomogram averaging) / Resolution: 30.0 Å
EMDB-4315, PDB-6ftg: Subtomogram average of OST-containing ribosome-translocon complexes from canine rough microsomal membranes Method: EM (subtomogram averaging) / Resolution: 9.1 Å
EMDB-4316, PDB-6fti: Cryo-EM Structure of the Mammalian Oligosaccharyltransferase Bound to Sec61 and the Programmed 80S Ribosome Method: EM (single particle) / Resolution: 4.2 Å
EMDB-4317, PDB-6ftj: Cryo-EM Structure of the Mammalian Oligosaccharyltransferase Bound to Sec61 and the Non-programmed 80S Ribosome Method: EM (single particle) / Resolution: 4.7 Å
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Keywords
PROTEIN TRANSPORT / Protein translocon of the endoplasmic reticulum
