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Amylopectin biosynthetic enzymes from developing rice seed form enzymatically active protein complexes.

Crofts N, Abe N, Oitome NF, Matsushima R, Hayashi M, Tetlow IJ, Emes MJ, Nakamura Y, Fujita N - J. Exp. Bot. (2015)

Bottom Line: This study investigated whether protein-protein interactions are also found in rice endosperm, as well as exploring differences between species.Blue-native-PAGE zymogram analyses confirmed the glucan-synthesizing activity of protein complexes.These results suggest that some rice starch biosynthetic isozymes are physically associated with each other and form active protein complexes.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Akita Prefectural University, 241-438 Kaidobata-Nishi, Shimoshinjo-Nakano, Akita city, Akita 010-0195, Japan.

No MeSH data available.


Related in: MedlinePlus

Possible protein–protein interactions in rice developing endosperm. Potential protein–protein interactions among starch biosynthetic enzymes of developing rice endosperm were deduced from western blotting (Fig. 2) and native-PAGE zymograms (Fig. 3) following GPC, co-immunoprecipitation experiments (Fig. 4), and BN-PAGE (Figs 5, 6) performed in this study. SS isozymes are in red, BE isozymes are in blue, DBE isozymes are in green, and Pho1 is in yellow. SSIIa is inactive in japonica rice (therefore not detected in Fig. 3) and indicated with white font. Single and double asterisks indicate formation of Pho1 dimers and ISA homo-oligomers confirmed in this study, previously reported by Hwang et al. (2010) and Utsumi et al. (2006), respectively. Other SS, BE, and DBE oligomers may occur, but are not included in this figure. The stoichiometric relationships between isozymes in high molecular weight complexes are unknown.
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Figure 7: Possible protein–protein interactions in rice developing endosperm. Potential protein–protein interactions among starch biosynthetic enzymes of developing rice endosperm were deduced from western blotting (Fig. 2) and native-PAGE zymograms (Fig. 3) following GPC, co-immunoprecipitation experiments (Fig. 4), and BN-PAGE (Figs 5, 6) performed in this study. SS isozymes are in red, BE isozymes are in blue, DBE isozymes are in green, and Pho1 is in yellow. SSIIa is inactive in japonica rice (therefore not detected in Fig. 3) and indicated with white font. Single and double asterisks indicate formation of Pho1 dimers and ISA homo-oligomers confirmed in this study, previously reported by Hwang et al. (2010) and Utsumi et al. (2006), respectively. Other SS, BE, and DBE oligomers may occur, but are not included in this figure. The stoichiometric relationships between isozymes in high molecular weight complexes are unknown.

Mentions: Based on GPC, co-immunoprecipitation, and BN-PAGE, possible combinations of protein–protein interactions in developing rice endosperm are postulated in Fig. 7. All of these protein complexes, in addition to the monomeric isozymes, may co-exist within the same cell in developing rice endosperm. The various dimeric interactions estimated at ~200kDa may be constituents of the larger protein complexes observed at 700kDa estimated to consist of SSIIa–SSIIIa–SSIVb–BEI, SSIIa–SSIIIa–BEIIb, SSIIa–SSIIIa–BEI–BEIIb–PUL, and SSI–SSIIa–SSIIIa–BEI–BEIIb–PUL. SSIIIa and BEIIa showed some interaction based on co-immunoprecipitation analyses, but the interaction was not detectable by western blotting after GPC or BN-PAGE. This suggests that the relative amounts of SSIIIa and BEIIb in this complex were low or possibly that the complex turns over very quickly and is lost during further electrophoretic and chromatographic separation.


Amylopectin biosynthetic enzymes from developing rice seed form enzymatically active protein complexes.

Crofts N, Abe N, Oitome NF, Matsushima R, Hayashi M, Tetlow IJ, Emes MJ, Nakamura Y, Fujita N - J. Exp. Bot. (2015)

Possible protein–protein interactions in rice developing endosperm. Potential protein–protein interactions among starch biosynthetic enzymes of developing rice endosperm were deduced from western blotting (Fig. 2) and native-PAGE zymograms (Fig. 3) following GPC, co-immunoprecipitation experiments (Fig. 4), and BN-PAGE (Figs 5, 6) performed in this study. SS isozymes are in red, BE isozymes are in blue, DBE isozymes are in green, and Pho1 is in yellow. SSIIa is inactive in japonica rice (therefore not detected in Fig. 3) and indicated with white font. Single and double asterisks indicate formation of Pho1 dimers and ISA homo-oligomers confirmed in this study, previously reported by Hwang et al. (2010) and Utsumi et al. (2006), respectively. Other SS, BE, and DBE oligomers may occur, but are not included in this figure. The stoichiometric relationships between isozymes in high molecular weight complexes are unknown.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4507757&req=5

Figure 7: Possible protein–protein interactions in rice developing endosperm. Potential protein–protein interactions among starch biosynthetic enzymes of developing rice endosperm were deduced from western blotting (Fig. 2) and native-PAGE zymograms (Fig. 3) following GPC, co-immunoprecipitation experiments (Fig. 4), and BN-PAGE (Figs 5, 6) performed in this study. SS isozymes are in red, BE isozymes are in blue, DBE isozymes are in green, and Pho1 is in yellow. SSIIa is inactive in japonica rice (therefore not detected in Fig. 3) and indicated with white font. Single and double asterisks indicate formation of Pho1 dimers and ISA homo-oligomers confirmed in this study, previously reported by Hwang et al. (2010) and Utsumi et al. (2006), respectively. Other SS, BE, and DBE oligomers may occur, but are not included in this figure. The stoichiometric relationships between isozymes in high molecular weight complexes are unknown.
Mentions: Based on GPC, co-immunoprecipitation, and BN-PAGE, possible combinations of protein–protein interactions in developing rice endosperm are postulated in Fig. 7. All of these protein complexes, in addition to the monomeric isozymes, may co-exist within the same cell in developing rice endosperm. The various dimeric interactions estimated at ~200kDa may be constituents of the larger protein complexes observed at 700kDa estimated to consist of SSIIa–SSIIIa–SSIVb–BEI, SSIIa–SSIIIa–BEIIb, SSIIa–SSIIIa–BEI–BEIIb–PUL, and SSI–SSIIa–SSIIIa–BEI–BEIIb–PUL. SSIIIa and BEIIa showed some interaction based on co-immunoprecipitation analyses, but the interaction was not detectable by western blotting after GPC or BN-PAGE. This suggests that the relative amounts of SSIIIa and BEIIb in this complex were low or possibly that the complex turns over very quickly and is lost during further electrophoretic and chromatographic separation.

Bottom Line: This study investigated whether protein-protein interactions are also found in rice endosperm, as well as exploring differences between species.Blue-native-PAGE zymogram analyses confirmed the glucan-synthesizing activity of protein complexes.These results suggest that some rice starch biosynthetic isozymes are physically associated with each other and form active protein complexes.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Akita Prefectural University, 241-438 Kaidobata-Nishi, Shimoshinjo-Nakano, Akita city, Akita 010-0195, Japan.

No MeSH data available.


Related in: MedlinePlus