Biosynthesis of alpha-Amylase in Vigna mungo Cotyledon.

In vitro translation of RNA extracted from Vigna mungo cotyledons showed that alpha-amylase is synthesized as a polypeptide with a molecular mass of 45,000, while cotyledons contain a form of alpha-amylase with a molecular mass of 43,000. To find out whether the 45,000 molecular mass polypeptide is a precursor to the 43,000 found in vivo, the cell free translation systems were supplemented with canine microsomal membrane; when mRNA was translated in the wheat germ system supplemented with canine microsomes, the 45,000 molecular mass form was not processed to a smaller form but the precursor form was partly processed in the membrane-supplemented reticulocyte lysate system. When V. mungo RNA was translated in Xenopus oocyte system, only the smaller form (molecular mass 43,000) was detected. Involvement of contranslational glycosylation in the maturating process of the alpha-amylase was ruled out because there was no effect of tunicamycin, and the polypeptide was resistant to endo-beta-H or endo-beta-D digestion. We interpret these results to mean that the 45,000 molecular mass form is a precursor with a signal peptide or transit sequence, and that the 43,000 molecular mass is the mature form of the protein.