The biosynthesis and processing of high molecular weight precursors of soybean glycinin subunits.
The predominant storage protein of soybean seed, glycinin, is composed of two heterogeneous classes of related subunits, the acidics (Mr approximately 38,000) and the basics (Mr approximately 22,000). Immunoreaction of polypeptides translated in vitro from isolated seed mRNA using antibodies prepared against either purified acidic or basic subunit groups precipitated precursor polypeptides of Mr = 60,000 to Mr = 63,000. High pressure liquid chromatography fingerprinting of trypsin-generated fragments from in vitro synthesized precursors showed fragments specific to both acidic and basic subunits. No mature acidic or basic subunits were detected in vitro translation reactions by either immunoprecipitation or high pressure liquid chromatography fingerprinting. Pulse-labeling of cotyledons growing in culture with [3H]glycine showed rapid accumulation of label in glycinin precursors of Mr = 59,000 to Mr = 62,000. Although in vivo synthesized precursors had slightly greater electrophoretic mobility than in vitro synthesized precursors, little label initially appeared in mature glycinin subunits. After several hours of continued cotyledon growth in absence of label, precursors were processed and label accumulated in both acidic and basic subunit groups. Recombinant plasmids were prepared by reverse transcription of soybean seed mRNA, and clones which encode glycinin precursors were identified by heteroduplex-hybridization of translatable messages. Northern blot analysis of seed mRNA shows the mRNA-encoding glycinin precursors to migrate at Mr = 0.71 X 10(6) on agarose gels, corresponding to approximately 2050 nucleotides. This is sufficiently large to encode a polypeptide consisting of both a glycinin acidic and basic subunit.
D002851:Chromatography, High Pressure Liquid; D004247:DNA; D005916:Globulins / Q000235:genetics*; D046911:Macromolecular Substances; D009693:Nucleic Acid Hybridization; D010446:Peptide Fragments / Q000032:analysis; D014674:Plant Proteins, Dietary / Q000235:genetics*; D010944:Plants / Q000378:metabolism; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D030262:Soybean Proteins; D013025:Glycine max
D005916:Globulins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D014674:Plant Proteins, Dietary; D012333:RNA, Messenger; D030262:Soybean Proteins; D012313:RNA; D004247:DNA; C011474:glycinin