Fig. 3

NcACBP functions as an active acyl-CoA-binding protein in vitro and in vivo. a Acyl-CoA binding activity of NcACBP. The SDS-PAGE analysis of purified recombinant protein NcACBP-GST from E. coli (a-i). Acyl-CoA binding activity of NcACBP was confirmed by incubating of the fluorescent substrate NBD-palmitoyl-CoA (0.25 μM) with GST-NcACBP (a-ii). b The binding kinetics of GST-NcACBP with NBD-palmitoyl-CoA were determined via a fluorescence assay. c Schematic illustrating the genetic complementation of ΔScACBP mutant yeast. d NcACBP gene (288 bp) can be detected in ΔScACBP:NcACBP clones. Lane 1 and lane 2 were different clones of ΔScACBP:NcACBP; lane 3 was ΔScACBP mutant yeast. e Phenotype rescue in yeast. Representative micrographs of wild-type S. cerevisiae yeast, ΔScACBP mutant yeast and ΔScACBP:NcACBP yeast were shown. Yeasts with single- and multilobed vacuoles were shown by yellow and blue arrowheads, respectively. f Yeast cells were categorized as normal or multilobed based on the phenotype. The bars indicate the means ± SD