Effect of structural modifications on the assembly of a glycinin subunit.
A Gy4 glycinin cDNA was modified and used to produce structurally altered 11S storage protein subunits. We evaluated these modified subunits for their ability to assemble into oligomers. Alterations made in the acidic polypeptide changed the subunit solubility characteristics but did not eliminate assembly. Modifications in the basic polypeptide usually eliminated assembly of subunits into trimers. A region exhibiting high natural variability located at the COOH terminus of the acidic polypeptide that we have designated the hypervariable region was also studied. Extensive deletions and insertions were tolerated in the hypervariable region without perturbing subunit assembly. Some of the insertions significantly increased the methionine content in the Gy4 glycinin subunit. Together, our results indicated that the structure of the basic polypeptide was more critical for assembly of trimers than that of the acidic polypeptide, an observation that implies that the basic polypeptides direct trimer formation. The assembly assays described here will be useful in efforts to improve seed quality. Using them, the effects of modifications to the storage protein subunits can be rapidly evaluated before introducing the mutated genes into plants.
Dickinson
C D
CD
United States Department of Agriculture, Purdue University, West Lafayette, Indiana 47907.
Scott
M P
MP
Hussein
E H
EH
Argos
P
P
Nielsen
N C
NC
D001483:Base Sequence; D004252:DNA Mutational Analysis; D017343:Genes, Plant / Q000235:genetics*; D005916:Globulins / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances; D008954:Models, Biological; D008969:Molecular Sequence Data; D014674:Plant Proteins, Dietary / Q000235:genetics / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D017434:Protein Structure, Tertiary; D030262:Soybean Proteins; D013025:Glycine max / Q000235:genetics / Q000378:metabolism*
D005916:Globulins; D046911:Macromolecular Substances; D014674:Plant Proteins, Dietary; D011498:Protein Precursors; D030262:Soybean Proteins; C011474:glycinin