Abstract
| Original language | English |
|---|---|
| Journal | Phytochemistry |
| Volume | 71 |
| Issue number | 14-15 |
| Pages (from-to) | 1650-1656 |
| ISSN | 0031-9422 |
| DOIs | |
| Publication status | Published - 2010 |
Keywords
- Cereal grain proteome
- Two-dimensional gel electrophoresis
- Seed germination
- Redox regulation
- Hordeum vulgare subsp vulgare
- Enzyme assays
Cite this
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Proteomic and activity profiles of ascorbate-glutathione cycle enzymes in germinating barley embryo. / Bønsager, Birgit Christine; Shahpiri, Azar; Finnie, Christine; Svensson, Birte.
In: Phytochemistry, Vol. 71, No. 14-15, 2010, p. 1650-1656.Research output: Contribution to journal › Journal article › Research › peer-review
TY - JOUR
T1 - Proteomic and activity profiles of ascorbate-glutathione cycle enzymes in germinating barley embryo
AU - Bønsager, Birgit Christine
AU - Shahpiri, Azar
AU - Finnie, Christine
AU - Svensson, Birte
PY - 2010
Y1 - 2010
N2 - Enzymes involved in redox control are important during seed germination and seedling growth. Ascorbate-glutathione cycle enzymes in barley embryo extracts were monitored both by 2D-gel electrophoresis and activity measurements from 4 to 144 h post imbibition (PI). Strikingly different activity profiles were observed. No ascorbate peroxidase (APX) activity was present in mature seeds but activity was detected after 24 h PI and increased 14-fold up to 144 h PI. In contrast, dehydroascorbate reductase (DHAR) activity was present at 4 h PI and first decreased by 9-fold until 72 h PI followed by a 5-fold increase at 144 h PI. Glutathione reductase and monodehydroascorbate reductase activities were also detected at 4 h PI, and showed modest increases of 1.8- and 2.7-fold, respectively, by 144 h PI. The combination of functional analysis with the proteomics approach enabled correlation of the activity profiles and protein abundance. While gel spots containing APX showed intensity changes consistent with the activity profile from 0 to 72 h PI, DHAR spot intensities indicated that post-translational regulation may be responsible for the observed changes in activity. Transcript profiling, 20-western blotting and mass spectrometric characterization of multiple APX spots demonstrated the presence of APX1 and minor amounts of APX2.
AB - Enzymes involved in redox control are important during seed germination and seedling growth. Ascorbate-glutathione cycle enzymes in barley embryo extracts were monitored both by 2D-gel electrophoresis and activity measurements from 4 to 144 h post imbibition (PI). Strikingly different activity profiles were observed. No ascorbate peroxidase (APX) activity was present in mature seeds but activity was detected after 24 h PI and increased 14-fold up to 144 h PI. In contrast, dehydroascorbate reductase (DHAR) activity was present at 4 h PI and first decreased by 9-fold until 72 h PI followed by a 5-fold increase at 144 h PI. Glutathione reductase and monodehydroascorbate reductase activities were also detected at 4 h PI, and showed modest increases of 1.8- and 2.7-fold, respectively, by 144 h PI. The combination of functional analysis with the proteomics approach enabled correlation of the activity profiles and protein abundance. While gel spots containing APX showed intensity changes consistent with the activity profile from 0 to 72 h PI, DHAR spot intensities indicated that post-translational regulation may be responsible for the observed changes in activity. Transcript profiling, 20-western blotting and mass spectrometric characterization of multiple APX spots demonstrated the presence of APX1 and minor amounts of APX2.
KW - Cereal grain proteome
KW - Two-dimensional gel electrophoresis
KW - Seed germination
KW - Redox regulation
KW - Hordeum vulgare subsp vulgare
KW - Enzyme assays
U2 - 10.1016/j.phytochem.2010.06.024
DO - 10.1016/j.phytochem.2010.06.024
M3 - Journal article
VL - 71
SP - 1650
EP - 1656
JO - Phytochemistry
JF - Phytochemistry
SN - 0031-9422
IS - 14-15
ER -