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. Author manuscript; available in PMC: 2018 Feb 26.
Published in final edited form as: Cell Rep. 2018 Feb 6;22(6):1365–1373. doi: 10.1016/j.celrep.2018.01.030

Figure 3. CPT1A Lysine Succinylates Enolase 1 to Inhibit Its Enzymatic Activity.

Figure 3

(A) Immunoprecipitated Flag-tagged enolase 1 was western blotted with anti-LVxx(succ)K succinylated lysine motif and anti-Flag antibodies. Lysates were western blotted for CPT1A and β-actin.

(B) Left: WB of the in vitro succinyltransferase assay samples using purified CPT1A WT or H473A and enolase 1 with anti-pan-succinylated lysine, anti-CPT1A, and anti-enolase 1 antibodies. Right: enolase activity of the in vitro succinyltransferase assay samples.

(C) WB of the in vitro succinyltransferase assay samples using purified CPT1A WT and BSA with anti-pan-succinylated lysine, anti-CPT1A, and anti-BSA antibodies.

(D) WB of the in vitro succinyltransferase assay samples at the indicated time points with anti-pan-succinylated lysine, anti-CPT1A, and anti-enolase 1 antibodies.

(E) WB of the in vitro succinyltransferase assay using the indicated amounts of purified CPT1A WT and enolase 1 with anti-pan-succinylated lysine, anti-CPT1A, and anti-enolase 1 antibodies.

(F) Enolase activity of the in vitro succinyltransferase assay samples using purified CPT1A WT with enolase 1 WT or 3KR.

Error bars, ±SD of three independent measurements. P values were determined using a two-tailed Student’s t test. **p < 0.01; n.s., not significant. See also Figure S3.