Human intestinal acyl-CoA synthetase 5 is sensitive to the inhibitor triacsin C

Figure 1 Sensitivity of recombinant acyl-CoA synthetase 5 proteins to triacsin C. Enzyme activity of Ni2+-affinity purified recombinant r-ACSL1, r-ACSL5, hu-ACSL5 C-terminal 6xHis-tagged enzyme, and hu-ACSL5 N-terminal 6xHis-tagged enzyme were assayed at pH 7.5 (white symbols) and pH 9.5 (black symbols) in the presence and absence of triacsin C as described in the methods section. ACSL5: Acyl-CoA synthetase 5.

Figure 2 Human acyl-CoA synthetase 5 is triacsin C sensitive in CaCo2 cells. A: CaCo2 cells were stably transfected with the full-length ACSL5 expressing pcDNA_DEST40 plasmid or empty control vector. Expression of N-terminal 6xHis-tagged hu-ACSL5 protein in CaCo2 was analyzed by Western blot using anti-ACSL5 and anti-His antibodies; B: Indirect immunofluorescence of N-terminal 6xHis-tagged hu-ACSL5 proteins, mitochondria, and nuclei in CaCo2 cells shows the mitochondrial localization of recombinant ACSL5 proteins. Original magnification × 400; C: Analysis of ACSL activity (pH 5-10) in hu-ACSL5 CaCo2 transfectants and controls (pcDNA_DEST40 CaCo2 transfectants and CaCo2 wild type) in the presence (white symbols) or absence (black symbols) of 25 μmol/L triacsin C. ACSL5: Acyl-CoA synthetase 5.

Figure 3 Acyl-CoA synthetase 5 related acyl-CoA synthetase activity is sensitive to triacsin C in human small intestinal mucosa. A: Human small intestinal mucosa after indirect immunofluorescence with anti-ACSL5 antibodies; B: Analysis of ACSL activity in human intestinal mucosa in the presence or absence of 25 μmol/L triacsin C. Triacsin C strongly inhibits ACSL activity in human intestinal mucosa. ACSL5: Acyl-CoA synthetase 5.