Table III.
Identification and relative carbonyl content of oxidized proteins during Arabidopsis seed aging
| No.a | Experimental Molecular Mass | Experimental pI | Arabidopsis Protein Name | Theoretical Molecular Mass | Theoretical pI | AGI No. | Coverage | Relative Abundanceb |
|---|---|---|---|---|---|---|---|---|
| kD | kD | ratio 7 d:0 d | ||||||
| Protein metabolism | ||||||||
| 140c | 60.26 | 4.15 | Calreticulin 1 (precursor) | 48.51 | 4.2 | At1g56340 | 15% | 2.3 |
| 1c | 79.36 | 5.08 | dnaK-type molecular chaperone BiP | 71.17 | 5.08 | At5g42020 | 20% | >100 |
| 135c | 80.5 | 5.04 | dnaK-type molecular chaperone BiP | 71.17 | 5.08 | At5g42020 | 20% | >100 |
| 136c | 77.87 | 4.87 | Heat shock cognate 70-kD protein 1 | 71.37 | 5.03 | At5g02500 | 19% | >100 |
| 43c | 78.9 | 5.06 | Heat shock cognate 70-kD protein 3 | 71.13 | 4.69 | At3g09440 | 15% | >100 |
| 137c | 76.06 | 5.07 | Heat shock protein 70 | 71.37 | 5.03 | At3g12580 | 38% | >100 |
| 614d | 68.68 | 5.83 | Aspartyl-tRNA synthetase | 62.90 | 6.12 | At4g31180 | 26% | 3.8 |
| 90c | 64.11 | 4.81 | Protein disulfide isomerase | 55.60 | 4.81 | At1g21750 | 39% | 2.1 |
| Energy metabolism and photosynthesis | ||||||||
| 109c | 64.79 | 6.35 | Malate oxidoreductase or malic enzyme | 64.28 | 6.32 | At2g19900 | 14% | >100 |
| 63c | 57.09 | 6.11 | Isocitrate lyase | 50.42 | 6.29 | At3g21720 | 30% | >100 |
| 10c | 57.36 | 5.77 | Ribulose bisphosphate carboxylase large chain | 53.47 | 6.25 | AtCg00490 | 30% | >100 |
| 319c | 57.36 | 5.57 | Ribulose bisphosphate carboxylase large chain | 53.47 | 6.25 | AtCg00490 | 26% | >100 |
| 320c | 57.36 | 5.82 | Ribulose bisphosphate carboxylase large chain | 53.47 | 6.25 | AtCg00490 | 21% | >100 |
| 321c | 57.36 | 5.84 | Ribulose bisphosphate carboxylase large chain | 53.47 | 6.25 | AtCg00490 | 30% | >100 |
| Stress response | ||||||||
| 60c | 66.68 | 5.74 | Late embryogenesis abundant (LEA) | 67.19 | 5.78 | At2g42560 | 25% | 1.2 |
| 61c | 66.68 | 5.78 | Late embryogenesis abundant (LEA) | 67.19 | 5.78 | At2g42560 | 43% | 1.5 |
| 431c | 66.68 | 5.79 | Late embryogenesis abundant (LEA) | 67.19 | 5.78 | At2g42560 | 32% | 2.2 |
| Hydrolase | ||||||||
| 96c | 64.62 | 6.08 | Glycosyl hydrolase family 1 protein | 57.83 | 6.02 | At3g21370 | 11% | >100 |
| 207c | 64.96 | 5.92 | Glycosyl hydrolase family 1 protein | 57.83 | 6.02 | At3g21370 | 21% | >100 |
| 208c | 64.96 | 6.23 | Glycosyl hydrolase family 1 protein | 57.83 | 6.02 | At3g21370 | 14% | >100 |
| Seed storage proteins | ||||||||
| 35c | 56.47 | 6.84 | 12S seed storage protein (precursor) | 55.86 | 6.36 | At4g28520 | 16% | >100 |
| 70c | 49.43 | 7.19 | 12S seed storage protein (precursor) | 52.59 | 7.68 | At5g44120 | 22% | >100 |
| 71c | 50.44 | 7.67 | 12S seed storage protein (precursor) | 52.59 | 7.68 | At5g44120 | 34% | >100 |
| 110c | 57 | 6.4 | 12S seed storage protein (precursor) | 55.86 | 6.36 | At4g28520 | 19% | >100 |
| 111c | 56.89 | 7.19 | 12S seed storage protein (precursor) | 55.86 | 6.36 | At4g28520 | 16% | >100 |
| 271d | 59.96 | 6.52 | 12S seed storage protein (precursor) | 55.86 | 6.36 | At4g28520 | 30% | >100 |
| 239c | 28.95 | 6.64 | α-Cruciferin 12S (seed storage protein fragment) | 48.03 | 6.56 | At1g03880 | 18% | >100 |
| 80c | 32.64 | 5.85 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 33% | 3.29 |
| 82c | 33.89 | 6.24 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 42% | 1.6 |
| 83c | 34.35 | 6.42 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 33% | 1 |
| 84c | 30.46 | 6.61 | α-Cruciferin 12S (seed storage protein fragment) | 31.75 | 6.49 | At5g44120 | 42% | 2.1 |
| 85c | 27.2 | 6.5 | α-Cruciferin 12S (seed storage protein fragment) | 27.24 | 6.34 | At1g03880 | 32% | 2.9 |
| 118c | 29.21 | 6.04 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 26% | 1 |
| 119c | 30.66 | 6.16 | α-Cruciferin 12S (seed storage protein fragment) | 31.75 | 6.49 | At5g44120 | 28% | 2.9 |
| 134c | 26.35 | 6.34 | α-Cruciferin 12S (seed storage protein fragment) | 27.24 | 6.34 | At1g03880 | 21% | 3.9 |
| 238c | 29.42 | 6.5 | α-Cruciferin 12S (seed storage protein fragment) | 31.75 | 6.49 | At5g44120 | 28% | 2.1 |
| 240c | 27.28 | 6.74 | α-Cruciferin 12S (seed storage protein fragment) | 27.24 | 6.34 | At1g03880 | 16% | 3.57 |
| 241c | 34.11 | 5.87 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 32% | 1.2 |
| 243c | 34.92 | 6.62 | α-Cruciferin 12S (seed storage protein fragment) | 34.68 | 6.42 | At4g28520 | 19% | 1 |
| 278c | 27.24 | 6.44 | α-Cruciferin 12S (seed storage protein fragment) | 31.75 | 6.49 | At5g44120 | 24% | >100 |
| 98c | 18.08 | 6.12 | β-Cruciferin 12S (seed storage protein fragment) | 20.8 | 7.03 | At1g03880 | 45% | 4.9 |
| 498c | 22.82 | 8.85 | β-Cruciferin 12S (seed storage protein fragment) | 21.2 | 6.19 | At4g28520 | 79% | 12.4 |
| 87c | 18.43 | 6.36 | β-Cruciferin 12S (seed storage protein fragment) | 20.8 | 7.03 | At1g03880 | 33% | >100 |
| 88c | 22.82 | 8.68 | β-Cruciferin 12S (seed storage protein fragment) | 21.2 | 6.19 | At4g28520 | 44% | 1.8 |
| 231c | 19.71 | 8.82 | β-Cruciferin 12S (seed storage protein fragment) | 21.2 | 6.19 | At4g28520 | 39% | >100 |
| 233c | 18.45 | 9.1 | β-Cruciferin 12S (seed storage protein fragment) | 20.84 | 9.06 | At5g44120 | 52% | >100 |
| 497c | 18.45 | 6.12 | β-Cruciferin 12S (seed storage protein fragment) | 21.2 | 6.19 | At4g28520 | 64% | >100 |
| 499c | 20.67 | 8.85 | β-Cruciferin 12S (seed storage protein fragment) | 20.84 | 9.06 | At5g44120 | 53% | 5.3 |
| Other processes | ||||||||
| 259c | 27.01 | 6.1 | Expressed protein | 27.28 | 6.7 | At1g05510 | 21% | 5.7 |
Protein numbering following Arabidopsis seed protein reference maps available at http://www.seed-proteome.com.
Data obtained from densitometric analysis of individual spots from carbonylated proteins on 2D gels revealed by anti-DNP immunoassay (examples are shown in Fig. 4): normalized spot volume in the deteriorated seeds (7 d of CDT) divided by the normalized spot volume in the nondeteriorated control seeds (0 d of CDT; ratio of carbonylation in deteriorated dry seeds [7 d of CDT] over carbonylation in nondeteriorated dry seeds [0 d of CDT]) from three different and independent protein extractions. >100 means that the accumulation level of the corresponding carbonylated protein in the nondeteriorated dry seeds (0 d of CDT) was close to background.
Listed proteins correspond to previously identified proteins (Gallardo et al., 2001, 2002a; Rajjou et al., 2004, 2006a; Job et al., 2005).
Listed proteins correspond to proteins identified during this work; the peptide sequences determined are available in Supplemental Table S2.