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. Author manuscript; available in PMC: 2022 Jul 1.
Published in final edited form as: Mol Cell. 2021 May 7;81(13):2693–2704.e12. doi: 10.1016/j.molcel.2021.04.013

Figure 5. WNK1 phenocopies EMC assembly to prevent promiscuous interactions of EMC2.

Figure 5.

(A) A complex of calmodulin (CaM) and 35S-methionine-labeled SEC61β (a tail-anchored (TA) protein that contains an exposed transmembrane domain (TMD)), containing the photocrosslinker BpA in the TMD, was generated in the PURE in vitro translation system. The CaM-SEC61β complexes were incubated with either purified EMC2 alone or complexes of EMC2 with the indicated binding partners. After UV irradiation the samples were analyzed by SDS-PAGE and autoradiography. (B) Quantification of the EMC2-SEC61β crosslink intensity from (A) (n=3). (C) Using the same assay as in (A), the CaM-SEC61β complexes were incubated with EMC2 alone or pre-formed complexes of EMC2 with increasing concentrations of purified wild type (WT) or L2250K mutant (Mut) WNK1 helix.