Characterisation of the storage protein subunits synthesised in vitro by polyribosomes and RNA from developing pea (Pisum sativum L.). II. Vicilin

Polypeptide material has been immunoprecipitated by antivicilin antibodies from translation products of polyribosomes and poly(A)-rich RNA isolated from developing seeds of Pisum sativum in the wheat germ and reticulocyte lysate cell-free synthesis systems. Analysis of this material by SDS-PAGE shows it to consist of three bands, of molecular weights 70,000, 50,000 and 47,000. The in vitro vicilin polypeptides of 70,000 and 50,000 mol. wt. have been shown to be very similar to the 70,000 and 50,000 mol. wt. subunits of vicilin by specific immunoprecipitation, and behaviour on treatment with cyanogen bromide and trypsin. The 50,000 mol. wt. in vitro vicilin polypeptide contains no significant extra sequence compared to the 50,000 mol. wt. vicilin subunit. The 47,000 mol. wt. in vitro vicilin polypeptide has no corresponding subunit in vicilin from mature seeds, but a 47,000 mol. wt. subunit is present in vicilin isolated from developing seeds. Comparison of translation products from polysomes isolated from seeds at middle and late stages of development shows that synthesis of the 50,000 and 47,000 mol. wt., but not 70,000 mol. wt. polypeptides is very much reduced at late stages of development. These results are discussed with reference to the nature of the vicilin fraction.