Figure 4.

C-terminal point mutant of Arabidopsis cry1 retains light response and ROS biosynthetic capacity. Wild type (a) and E531K (b) mutant proteins were expressed and purified from baculovirus expression system as previously described³³. ‘Dark’ shows spectra of isolated proteins prior to illumination. ‘Light’ indicates spectra after photoreduction for 30 seconds at 500 μmol m⁻² s⁻¹ blue light in the presence of 5 mM DTT . Samples were subsequently returned to dark and spectra taken at the indicated times (in min). (c). 100micromolar concentration of proteins were illuminated at 500 μmol m⁻² s⁻¹ blue light and aliquots taken at the indicated time for detection of ROS by AMPLEX RED fluorescence detection method as described¹⁶. Aliquots were taken at the indicated times. Error bars represent SD of three independent measurements.