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. Author manuscript; available in PMC: 2019 Jul 20.
Published in final edited form as: ACS Chem Biol. 2018 Feb 8;13(7):1824–1831. doi: 10.1021/acschembio.7b00925

Figure 1.

Figure 1.

C117S is a conserved sensor of HNE. (A) Schematic for T-REX in living systems. Also see Figure S1 in the Supporting Information. (B) Homology models of (left) zebrafish Hspb7 (surface-accessible and ribbon) and (right) human HSPB7 (ribbon). Note that, for the surface-accessible model, sulfur within C117 is colored red. (C) Phylogenetic analysis of HSPB7s. Clade colored red contains C49. Also see Figures S2 and S3 in the Supporting Information. (D) Sequence logos for C117 and C49. (E) HEK293T cells were transfected with respective plasmids encoding Halo-HSPB7(wt/mutants), grown for 24 h, then treated with Ht-PreHNE (20 μM, 2 h), washed and exposed to light. Cells were lysed and HNEylated-proteins were labeled with Cy5-azide; see Figure S1 for workflow. Lysates were resolved by SDS-PAGE and imaged by Cy5, and blotted for the stated proteins. (F) Quantitation of data in panel (E).