In vitro synthesis of the Bowman-Birk and related soybean protease inhibitors

Poly (A)(+) RNAs from immature soybean seeds were size fractionated in denaturing sucrose gradients to identify mRNA that directs the cell-free synthesis of the Bowman-Birk protease inhibitor and the related inhibitors PI I-IV. Polypeptides synthesized in vitro were labeled with ((35)S)-cysteine and ((3)H)-serine and detected by immunoprecipitation with anti Bowman-Birk and anti PI I-IV sera. Immunoprecipitates of the translation products comigrated on SDS-polyacrylamide gels with the dimeric or trimeric aggregates of the authentic inhibitor proteins, which self-associate under certain conditions. Further evidence that these immunoprecipitates contained authentic polypeptides corresponding to the Bowman-Birk or PI IV inhibitor was shown by sequential amino acid analyses of peptides generated by cleavage with cyanogen bromide.