Figure 6.
Expression levels and enzyme activity of hPanK1, hPanK2mtmΔ and hPanK3 in yeast. (A) Western blot analysis using anti-Myc monoclonal antibody on extracts from the yeast mutant strains carrying human PANK1, PANK2mtmΔ and PANK3. (B) Quantification of the expression levels of human PanKs in yeast from (A) using Image Lab software. The signals were normalized to the control signal (α-Por1). Values are means ± standard deviation, n = 3. (C,D) Cellular pantothenate kinase was measured from fresh cell extracts obtained from cab1Δ/cab1G351S yeast cells carrying either the empty vector, yeast CAB1 or human PANK genes. Cells were grown at 30 °C in media containing 2% galactose. The activity assay measures the phosphorylation of D-[1-14C] pantothenate as described in Material and Methods.