Reconstitution of Single Molecular Species from Isolated Subunits of Glycinin

摘要

Ion-exchange HPLC systems using POROS 20 HQ and Mono Q HR 10/10 columns were applied to isolate glycinin subunits under deaturing conditions. Analyses by SDS-PAGE, N-terminal amino acid sequence, and sucrose density gradient centrifugation showed that the pseudoglycinin from the highly purified homo-subunit, A_3B_4, was reconstituted. The A_3B_4 pseudoglycinin was similar to the native glycinin with respect to molecular size, subunit structure, and secondary structure. The hexameric pseudoglycinin dissociated into trimers after long stroage at pH 7.6.