Table 2. The identification of the differential extracted proteins in maize using the two methods.
| Protein name | Accession | MW(kDa)/pI | Protein score |
Coverage (%) |
GRAVY | Subcellular localization |
Matched Peptides |
|---|---|---|---|---|---|---|---|
| Calreticulin-2, partial (spot 1) | ONM21219 | 41.11/4.81 | 240 | 47.6 | -0.870 | Endoplasmic reticulum | FEDGWESR;NPNYQGKWK;QTGSIYEHWDILPPK;WKAPMIDNPDFK; FYAISAEYPEFSNK;YIGIELWQVK;KDDNMAGEWNHTSGK;SGTLFDNIIITDDPALAK;WNGDAEDKGIQTSEDYR;KPEGYDDIPKEIPDPDAK;KPEDWDDKEYIPDPEDK;KPEDWDDEEDGEWTAPTIPNPEYK; |
| Fructokinase-1 (spot 2) | AAP42805 | 34.67/4.87 | 86 | 39.0 | 0.139 | Chloroplast | MLAAILR;EALWPSR;EFMFYR;LGDDEFGR;DFHGAVPSFK;TAHLRAMEIAK;FANACGAITTTK;DNGVDDGGVVFDSGAR;LGGGAAFVGKLGDDEFGR;AAVFHYGSISLIAEPCR;VSEVELEFLTGIDSVEDDVVMK |
| Glycine-rich RNA-binding protein 2(spot 3) | ACG28116 | 15.48/6.1 | 182 | 34.6 | -0.487 | Nucleus | NITVNEAQSR;GGGYGNSDGNWR;RDGGGGYGGGGGGYGGGGGYGGGGGGYGGGNR |
| Elongation factor 1-β (spot 4) | ONM59608 | 15.67/4.38 | 134 | 13.2 | -0.360 | Cell membrane | LDEYLLTR;KLDEYLLTR;LSGITAEGQGVK;WFNHIDALVR |
| Glutathione transferase (spot 5) |
CAA73369 | 25.10/6.21 | 132 | 43.8 | -0.174 | Cytoplasm | GDGQAQAR;NLYPPEK;LYDCGTR;AEMVEILR;KLYDCGTR;VYDFVCGMK;FWADYVDKK;ECPRLAAWAK;GLAYEYEQDLGNK;QGLQLLDFWVSPFGQR;KQGLQLLDFWVSPGQR;QGLQLLDFWVSPFGQRCR;GLAYEYLEQDLGNKSELLLR |
| Lea14-A (spot 6) |
AMY96568 | 16.08/5.64 | 514 | 96.7 | 0.024 | Chloroplast. Golgi apparatus. Nucleus. | DGATLAGRVDVR;DWDIDYEMR;SGELKLPTLSSIF;DAGRDWDIDYEMR;LDVPVKVPYDFLVSLAK;TVASGTVPDPGSLAGDGATTR;VGLTVDLPVVGKLTLPLTK;NPYSHAIPVCEVTYTLR;LANIQKPEAELADVTVGHVGR;SAGRTVASGTVPDPGSLAGDGATTR;VDVRNPYSHAIPVCEVTYTLR;GFVADKLANIQKPEAELADVTVGHVGR |
| Sedoheptulose-1,7-bisphosphatase (spot 11) |
ACG31345 | 41.79/6.8 | 101 | 19.0 | -0.136 | Chloroplast | EKYTLR;LLICMGEAMR;FEETLYGSSR;GIFTNVTSPTAK;YTGGMVPDVNQIIVK;LTGVTGGDQVAAAMGIYGPR |
| UDP-glucose 6-dehydrogenase (spot 14) |
AQK62747 | 53.53/5.71 | 545 | 27.0 | -0.072 | Chloroplast | LAANAFLAQR; NLFFSTDVEK; AADLTYWESAAR; AADLTYWESAAR; DVYAHWVPEDR; ILTTNLWSAELSK; CPDIEVVVVDISKPR; IFDNMQKPAFVFDGR; AQISIYDPQVTEDQIQR; GINYQILSNPEFLAEGTAIEDLFKPDR |
| peroxidase 39 precursor (spot 15) |
NP_001149755 | 35.81/7.59 | 364 | 27.0 | -0.002 | Cytoplasm | DAAPNLTLR; TFDLSYYR; THFHDCFVR; AHIPHAPDVASTLLR; GLFQSDAALITDAASK; DSVGVIGGPFWSVPTGR; ECPGVVSCADIVALAAR |
| proteasome subunit alpha type 1 (spot 17) |
NP_001149085 | 30.28/5.19 | 440 | 41.0 | -0.293 | Nucleus | LSSSNCTVAIVGR; VADHAGVALAGLTADGR; EDGTIEPFEMIGAAR; FQGYNDYTPEQLIK; NQYDTDVTTWSPQGR; KEDGTIEPFEMIGAAR; NQYDTDVTTWSPQGR; NECINHSFVYEAPLPVSR; SSTHAVLAAVNKPASELSSYQR |
| Cold shock protein 2 (spot 18) |
AQK60690 | 23.90/5.95 | 88 | 8.0 | -0.635 | Golgi apparatus; Nucleus. | GFGFISPEDGSEDLFVHQSSIK |
| 16.9 kDa class I heat shock (spot 21) |
NP_001146967 | 17.05/6.77 | 302 | 26.0 | -0.451 | Nucleus | FRLPENAK; AALENGVLTVTVPK; VEVEDGNVLLISGQR; EEVKVEVEDGNVLLISGQR |
| Profilin-1 (spot 25) |
AQL00684 | 9.69/5.14 | 54 | 36.0 | -0.172 | Chloroplast; Cytoplasm. | DFDEPGFLAPTGLFLGPTK |
| ABA-inducible gene protein (spot 26) |
CAA31077 | 15.49/5.55 | 368 | 52 | -0.718 | Nucleus | AAADVEYR; DGGYGGGGGYGGR; EGGGGGYGGGGGYGGR; EGGGGGYGGGGGGWRD; REGGGGGYGGGGGYGGR; GFGFVTFSSENSMLDAIENMNGK; GFGFVTFSSENSMLDAIENMNGKELDGR |
Note: For MS/MS analysis, differential abundance spots (>2 folds) were extracted, in-gel digested (trypsin, 37°C, 20 h), and analyzed by the MALDI-TOF/TOF analyzer (AB SCIEX TOF/TOF-5800, USA). MALDI-TOF/TOF spectra were acquired in the positive ion mode and automatically submitted to Mascot 2.2 (http://www.matrixscience.com) for identification against NCBInr database (version Sept 29, 2018; species, Zea mays, 719230 sequences). The search parameters were as follows: type of search: combined (MS + MS/MS); enzyme: trypsin; dynamical modifications: oxidation (M); fixed modifications: carbamidomethyl (C); mass values: monoisotopic; protein mass: unrestricted; peptide mass tolerance: ±100 ppm; fragment mass tolerance: ±0.4 Da; peptide charge state: 1+; max missed cleavages: 1. Unambiguous identification was judged by the number of matched peptide sequences, sequence coverage, Mascot score, and the quality of MS/MS spectra. All of the positive protein identification scores were significant (p<0.05).