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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

Starch biosynthetic enzyme activities analysed on non-denaturing zymograms and by western blotting following GPC of rice developing endosperm. Numbers at the top indicate the molecular weight of protein standards in kiloDaltons. (A) SS activity. (B) BE activity. (C) DBE activity. (D–G) Western blots of identical native-PAGE used for (A) were probed with SSI, SSIIa, SSIIIa, and SSIVb antibodies, respectively. (H–K) Western blots of identical native-PAGE used for (B) were probed with BEI, BEIIa, BEIIb, and Pho1 antibodies, respectively. (L) Western blots of identical native-PAGE used for (C) were probed with PUL antibody. Black arrowheads in (A–C) indicate activities of isozymes. Grey arrowheads in (A) indicate glycosyl hydrolase or glucan transferase activities. Arrowheads in (D–L) indicate polypeptides recognized by the antibody.
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Figure 3: Starch biosynthetic enzyme activities analysed on non-denaturing zymograms and by western blotting following GPC of rice developing endosperm. Numbers at the top indicate the molecular weight of protein standards in kiloDaltons. (A) SS activity. (B) BE activity. (C) DBE activity. (D–G) Western blots of identical native-PAGE used for (A) were probed with SSI, SSIIa, SSIIIa, and SSIVb antibodies, respectively. (H–K) Western blots of identical native-PAGE used for (B) were probed with BEI, BEIIa, BEIIb, and Pho1 antibodies, respectively. (L) Western blots of identical native-PAGE used for (C) were probed with PUL antibody. Black arrowheads in (A–C) indicate activities of isozymes. Grey arrowheads in (A) indicate glycosyl hydrolase or glucan transferase activities. Arrowheads in (D–L) indicate polypeptides recognized by the antibody.

Mentions: The same GPC fractions were analysed by native-PAGE/activity staining (Fig. 3A–C). The fractions with the highest enzymatic activity (Fig. 3) generally correlated with the strongest signals obtained by SDS–PAGE and western blotting (Fig. 2).


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)

Starch biosynthetic enzyme activities analysed on non-denaturing zymograms and by western blotting following GPC of rice developing endosperm. Numbers at the top indicate the molecular weight of protein standards in kiloDaltons. (A) SS activity. (B) BE activity. (C) DBE activity. (D–G) Western blots of identical native-PAGE used for (A) were probed with SSI, SSIIa, SSIIIa, and SSIVb antibodies, respectively. (H–K) Western blots of identical native-PAGE used for (B) were probed with BEI, BEIIa, BEIIb, and Pho1 antibodies, respectively. (L) Western blots of identical native-PAGE used for (C) were probed with PUL antibody. Black arrowheads in (A–C) indicate activities of isozymes. Grey arrowheads in (A) indicate glycosyl hydrolase or glucan transferase activities. Arrowheads in (D–L) indicate polypeptides recognized by the antibody.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 3: Starch biosynthetic enzyme activities analysed on non-denaturing zymograms and by western blotting following GPC of rice developing endosperm. Numbers at the top indicate the molecular weight of protein standards in kiloDaltons. (A) SS activity. (B) BE activity. (C) DBE activity. (D–G) Western blots of identical native-PAGE used for (A) were probed with SSI, SSIIa, SSIIIa, and SSIVb antibodies, respectively. (H–K) Western blots of identical native-PAGE used for (B) were probed with BEI, BEIIa, BEIIb, and Pho1 antibodies, respectively. (L) Western blots of identical native-PAGE used for (C) were probed with PUL antibody. Black arrowheads in (A–C) indicate activities of isozymes. Grey arrowheads in (A) indicate glycosyl hydrolase or glucan transferase activities. Arrowheads in (D–L) indicate polypeptides recognized by the antibody.
Mentions: The same GPC fractions were analysed by native-PAGE/activity staining (Fig. 3A–C). The fractions with the highest enzymatic activity (Fig. 3) generally correlated with the strongest signals obtained by SDS–PAGE and western blotting (Fig. 2).

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