{alpha}' Subunit of soybean {beta}-conglycinin forms complex with rice glutelin via a disulphide bond in transgenic rice seeds.
The alpha' and beta subunits of soybean beta-conglycinin were expressed in rice seeds in order to improve the nutritional and physiological properties of rice as a food. The alpha' subunit accumulated in rice seeds at a higher level than the beta subunit, but no detectable difference in mRNA transcription level between subunits was observed. Sequential extraction results indicate that the alpha' subunit formed one or more disulphide bonds with glutelin. Electron microscopic analysis showed that the alpha' subunit and the beta subunit were transported to PB-II together with glutelin. In mature transgenic seeds, the beta subunit accumulated in low electron density regions in the periphery of PB-II, whereas the alpha' subunit accumulated together with glutelin in high-density regions of the periphery. The subcellular localization of mutated alpha' subunits lacking one cysteine residue in the N-terminal mature region (alpha'DeltaCys1) or five cysteine residues in the pro and N-terminal mature regions (alpha'DeltaCys5) were also examined. Low-density regions were formed in PB-II in mature seeds of transgenic rice expressing alpha'DeltaCys 5 and alpha'DeltaCys1. alpha'DeltaCys5 was localized only in the low-density regions, whereas alpha'DeltaCys1 was found in both low- and high-density regions. These results suggest that the alpha' subunit could make a complex via one or more disulphide bonds with glutelin and accumulate together in PB-II of transgenic rice seeds.
Motoyama
Takayasu
T
Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto, Japan 611-0011.
Maruyama
Nobuyuki
N
Amari
Yoshiki
Y
Kobayashi
Kanna
K
Washida
Haruhiko
H
Higasa
Takahiko
T
Takaiwa
Fumio
F
Utsumi
Shigeru
S
D052179:Antigens, Plant / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005916:Globulins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005983:Glutens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012275:Oryza / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055314:Seed Storage Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030262:Soybean Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013025:Glycine max / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D052179:Antigens, Plant; D005916:Globulins; D021122:Protein Subunits; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; D005983:Glutens