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. Author manuscript; available in PMC: 2017 Jul 20.
Published in final edited form as: Science. 2017 Jan 20;355(6322):298–302. doi: 10.1126/science.aah6130

Fig. 2. Reconstitution of TA protein triage with purified factors.

Fig. 2

(A) Recombinant TA-SGTA and TA-TRC40 complexes (fig. S5; TA protein contains the Vamp2 TMD) were assayed for insertion into ER-derived rough microsomes (RMs) by TA protein glycosylation (glyc). (B) TA protein (Vamp2) in complex with recombinant Bag6 or ∆Ubl-Bag6 was assayed for ubiquitination by RNF126. (C) TA-SGTA complex (Vamp2) was incubated with the indicated components and analyzed for insertion into RMs by glycosylation. The small amount of insertion seen in lane 4 is due to residual TRC40 that copurified with the RMs. (D) TA-SGTA complex (Vamp2) was incubated with the indicated components and assayed for TA protein ubiquitination. All reactions were conducted with 650 nM of each TRC component.

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