CHARACTERIZATION AND PURIFICATION OF THE FUSICOCCIN-BINDING COMPLEX FROM PLASMA-MEMBRANES OF COMMELINA-COMMUNIS

The fungal phytotoxin fusicoccin binds with high affinity to plasma membranes of the monocotyledonous plant, Commelina communis L. The sites bind the toxin with an apparent K(d) of 5.2 nM and a pH optimum of 6.0. They occur at a level of approximately 6-8 pmol/mg plasma membrane protein. Photoaffinity labeling with the biologically active fusicoccin derivative 9'-nor-8'-(4-azido[3,5-H-3]benzoyl) diaminoethylfusicoccin identified a polypeptide of 31.5 kDa on SDS/PAGE which was strongly labeled. A second 32.5-kDa band was also consistently labeled, although not to the same extent. The binding sites were solubilized in functional form and a purification scheme was developed based on affinity and ion-exchange procedures. The purified fraction contains two polypeptides of apparent molecular masses of 30.5 kDa and 31.6 kDa. A detailed molecular analysis of the fusicoccin-binding complex is now possible.