Expression in Escherichia coli of the two subunits of the isozyme form of wheat germ protein synthesis initiation factor 4F. Purification of the subunits and formation of an enzymatically active complex.

The subunits (p28 and p86) of the isoenzyme form of eukaryotic initiation factor 4F (eIF-(iso)4F) from wheat were expressed separately in Escherichia coli. The subunits were purified by affinity chromatography (p28) and ion-exchange chromatography (p86). The purified subunits alone did not support polypeptide synthesis in an eIF-(iso)4F and eIF-4F-deficient translation system from wheat germ. However, when the two subunits were mixed together, activity equal to that of the native form of eIF-(iso)4F was obtained. These results show that subunits expressed separately are able to associate and form an enzymatically active complex.