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TitleStructural insights into TRAP association with ribosome-Sec61 complex and translocon inhibition by a CADA derivative.
Journal, issue, pagesSci Adv, Vol. 9, Issue 9, Page eadf0797, Year 2023
Publish dateMar 3, 2023
AuthorsEva Pauwels / Neesha R Shewakramani / Brent De Wijngaert / Anita Camps / Becky Provinciael / Joren Stroobants / Kai-Uwe Kalies / Enno Hartmann / Piet Maes / Kurt Vermeire / Kalyan Das /
PubMed AbstractDuring cotranslational translocation, the signal peptide of a nascent chain binds Sec61 translocon to initiate protein transport through the endoplasmic reticulum (ER) membrane. Our cryo-electron ...During cotranslational translocation, the signal peptide of a nascent chain binds Sec61 translocon to initiate protein transport through the endoplasmic reticulum (ER) membrane. Our cryo-electron microscopy structure of ribosome-Sec61 shows binding of an ordered heterotetrameric translocon-associated protein (TRAP) complex, in which TRAP-γ is anchored at two adjacent positions of 28 ribosomal RNA and interacts with ribosomal protein L38 and Sec61α/γ. Four transmembrane helices (TMHs) of TRAP-γ cluster with one C-terminal helix of each α, β, and δ subunits. The seven TMH bundle helps position a crescent-shaped trimeric TRAP-α/β/δ core in the ER lumen, facing the Sec61 channel. Further, our in vitro assay establishes the cyclotriazadisulfonamide derivative CK147 as a translocon inhibitor. A structure of ribosome-Sec61-CK147 reveals CK147 binding the channel and interacting with the plug helix from the lumenal side. The CK147 resistance mutations surround the inhibitor. These structures help in understanding the TRAP functions and provide a new Sec61 site for designing translocon inhibitors.
External linksSci Adv / PubMed:36867692 / PubMed Central
MethodsEM (single particle)
Resolution2.79 - 2.86 Å
Structure data

15860

8b5l

EMDB-15860, PDB-8b5l:
Cryo-EM structure of ribosome-Sec61-TRAP (TRanslocon Associated Protein) translocon complex
Method: EM (single particle) / Resolution: 2.86 Å

15863

8b6c

EMDB-15863, PDB-8b6c:
Cryo-EM structure of ribosome-Sec61 in complex with cyclotriazadisulfonamide derivative CK147
Method: EM (single particle) / Resolution: 2.79 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-OWI:
4-[[9-(cyclohexylmethyl)-3-methylidene-5-(4-methylphenyl)sulfonyl-1,5,9-triazacyclododec-1-yl]sulfonyl]-~{N},~{N}-dimethyl-aniline

Source
  • Canis lupus familiaris (dog)
  • Rabbit (rabbit)
  • oryctolagus cuniculus (rabbit)
KeywordsMEMBRANE PROTEIN / Ribosome / Sec61 / TRAP / Translocon / Translation / RNA binding protein / eEF2 / CK147 / CADA / Translocon inhibitor / 80S / 60S / 40S / cyclotriazadisulfonamide

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