Proteasomes from Dictyostelium discoideum: characterization of structure and function.

We have isolated and purified 20 S proteasomes from Dictyostelium discoideum and characterized their proteolytic activities. Two-dimensional electrophoresis revealed 13 distinct spots. Affinity purification with a subunit-specific monoclonal antibody indicated that the preparation was homogeneous, i.e., each individual proteasome appeared to have the full set of subunits. We have not obtained any evidence for changes in subunit composition at different developmental stages. The cDNA clones coding for two subunits (4 and 5), both of the alpha-type, have been cloned and sequenced. It has been shown by immunoelectron microscopy that each proteasome is composed of two identical halves, related to each other by C2 symmetry. The resulting model implies that the alpha- and beta-subunits have a fixed pattern of relationships. D. discoideum proteasomes are found both in the cytosol and, in higher concentrations, in the nucleus.