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Table S1: Data collection and refinement statistics.

Figure S1: Cross-eyed stereo view of electron density for the Mg2+ADPAlF4complex of M. thermautotrophicus TRC40. The refined model is superimposed onto a σA-weighted 2FoFc map calculated at 2.1 Å resolution and contoured at 2σ.

Figure S2: Characterization of the depleted translation extract (PD-RRL). Sec61β was translated in PD-RRL in the presence or absence of recombinant yeast Get3. An aliquot was subjected to chemical cross-linking with DSS to evaluate substrate interactions with 5 ng/µL Get3 (top panel). The remainder was incubated with yRMs and analyzed for insertion by the protease protection assay (bottom panel). PD-RRL lacks all known chaperones that interact with ER-directed TA proteins, and is deficient for insertion; activity was restored by supplementing with physiological levels of TRC40/Get3.

TRA_1229_sm_suppinfo.pdf1565KSupporting info item

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