Fig. 5

Calcium/calmodulin binds to FvUGT1 in vitro. a. Co-precipitation of FvUGT1 with calmodulin-agarose beads in the presence of calcium. The total proteins (lysates) from E. coli cells carrying either pET-FvUGT1-His-tag or negative control (pET32) were applied to calmodulin-agarose beads in the presence of 1 mM CaCl₂ or 1 mM EGTA. Proteins bound to the beads were eluted and analyzed using Western blotting against an anti-His-tag antibody. b. Gel mobility shift assay showing that calcium/calmodulin binds the synthetic peptide corresponding to the putative calmodulin-binding region in FvUGT1. The synthetic peptide sequence is shown at the top of panel b. Arrows indicate the positions of free calmodulin and the calmodulin/peptide complex. CaM, calmodulin