Acyl-CoA oxidase ACOX-1 interacts with a peroxin PEX-5 to play roles in larval development of Haemonchus contortus
Fig 2
Enzyme activity and spotting assay of Hc-ACOX-1 in vivo and in vitro.
(A-C) Linear double-reciprocal plots of Hc-ACOX-1.1 (A), Hc-ACOX-1.2 (B) and Hc-ACOX-1.3 (C) constructed based on reciprocal reaction velocity (1/V) and reciprocal value of palmitoyl-CoA concentration (1/[S]). Panels D-G: The letters (1.1, 1.2 and 1.3) in square brackets represent the corresponding protein of Hc-ACOX-1 and the characters in parentheses indicate the key sites that are replaced with Alanine. 151A/190A represents the 151st and 190th amino acids are replaced with Alanine. Yeast concentrations are marked at top with a starting concentration of OD600 = 1. (D) Growth of wild-type Saccharomyces cerevisiae on YNBO plates (YNB supplemented with oleic acid as sole carbon source). (-) represents mutation without peroxisomal targeting signal type 1. (E) Growth of S. cerevisiae Δpox1 strain rescued with Hc-acox-1 on YNBO plates. (F) Effect of key site of Hc-ACOX-1.2 and -1.3 on the growth of Δpox1. (G) Effect of key sites of Hc-ACOX-1.1 on the growth of Δpox1. Multiple mutations in Hc-ACOX-1.1 affect the growth of Δpox1 on the YNBO plate, whereas single mutation at 151A or 190A did not. WT and Δpox1 represent wild-type and POX (ACOX homologue) mutant strain of S. cerevisiae, respectively.