Conformation and activity of Phaseolus coccineus var. rubronanus lectin.

The conformation of native and denatured Phaseolus coccineus var. rubronanus lectin was studied by circular dichroism (CD) and correlated to the hemagglutinating activity. The far-UV CD spectrum at 25 degrees C showed a broad, negative band around 223 nm and a positive one at 196 nm. CD data analysis of the lectin indicated a beta-sheet-rich protein. At high temperatures, the spectrum was blue-shifted with increasing magnitude; these changes correlated well with the loss of the activity. The conformation of lectin between pH 2 and 10 remained essentially unchanged. At pH 13 the CD spectrum resembled that of unordered form with a negative band near 200 nm and the activity was completely lost. The denatured lectin in 6 M guanidine hydrochloride would be renatured upon diluting the denaturant to 0.75 M; the changes in CD spectrum again correlated well with the loss of the activity. The effect of sodium dodecyl sulfate on the lectin was drastic; it sharply increased the alpha-helix at the expense of the beta-sheet and reduced the activity; the changes reached a plateau above 20 mM surfactant.