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. Author manuscript; available in PMC: 2019 May 18.
Published in final edited form as: ACS Chem Biol. 2018 Apr 6;13(5):1130–1136. doi: 10.1021/acschembio.7b01063

Figure 2. 2-HD covalently modifies the conserved Cys62 of Bax and induces Bax-dependent cell death.

Figure 2

(A) Probe 2 induces Bax-dependent cell death in HCT116 wild type cells, compared to HCT116 Bax/Bak DKO cells. (B) Quantification of cell death effects induced by Probe 2. Data are represented as mean +/− SD, n=3, **, P<0.01, ***, P<0.001. (C) Probe 2 induces Bax-dependent increase of ROS, while 4-HNE induce Bax-independent ROS. Data are represented as mean +/− SD, n=3, **, P<0.01. (D) Cys62 is the major site of 2-HD modification on Bax. Probe 2 partially labels Bax C62S and does not label C62/126S (2CS) mutant, while labeling wild type and C126S mutant efficiently. (E) Detection of the probe-labeled peptide containing Cys62 by mass spectrometry. Observed the m/z of 1054.635, which matches the probe-modified Cys62 in peptide KLSECLK.