Table I.
Identification of potential Trx targets in germinating M. truncatula seeds
The labeled and isolated targets identified in axis (A) or cotyledon (C) extracts showed an E-value <−3. Available tentative consensus (TC), EST, and accession numbers corresponding to M. truncatula proteins are mentioned. Proteins for which only partial sequences are known in M. truncatula, the accession numbers of the nearest matching orthologs, are indicated. Key properties are also listed: theoretical MW/pI, coverage (%), peptide number, conserved/total Cys, and potential localization. Predotar and Psort were used to determine potential localization of complete proteins known in M. truncatula or others species. C, Cytosol; M, mitochondria; ER, endoplasmic reticulum; CP, chloroplasts; N, nucleus. Proteins identified with only one peptide are indicated by §. The presence of the protein in the 0.2 m NaCl fraction is denoted with *. The three proteins that have no Cys residues (spots 63, 101, 108) were not further considered in the study.
| Spot No.
|
Protein
|
E-Value
|
Protein Labeled with mBBr
|
Protein Isolated on Mutant Trx
|
M. truncatula TC, EST, or Accession No.
|
Nearest Ortholog Accession No.
|
Theoretical MW/pI
|
Coverage (%)
|
Peptide No.
|
Conserved/Total Cys
|
Potential Localization
|
||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| A | C | A | C | ||||||||||
| 1 | UK1 | −15 | • | TC101829 | – | 15 | 3 | ||||||
| 2 | UK2 | −7 | • | TC97992 | – | 6.5 | 2 | ||||||
| 3 | HSP90 | −60 | • | TC94880 | 95.5/5.1 | 12 | 8 | 12/14 | C | ||||
| 4 | CDC48 | −147 | • | • | TC94474 | 89.7/5.1 | 32 | 17 | 13/15 | C | |||
| −117 | • | • | TC97683 | P54774 | 89.7/4.9 | 4.1 | 14 | 14/14 | C | ||||
| 5 | ADP-ribosyltransferase or PM38 | −92 | • | • | BQ124797 | ADD51626 | 91.6/5.2 | 15 | 11 | 18/20 | C | ||
| 6 | EF2 | −181 | • | • | • | TC93936 | 94.1/5.8 | 39 | 21 | 15/16 | C | ||
| −14 | • | TC94039 | Xp_465992 | 94.0/5.7 | 3.7 | 3 | 15/18 | C | |||||
| −72 | • | • | TC94015 | 94.1/5.8 | 7.2 | 10 | 15/17 | C | |||||
| 7 | Lipoxygenase | −270 | • | • | TC94816 | P09439 | 97.1/6.2 | 49 | 28 | 4/5 | C | ||
| 8 | HSP90 (putative)§ | −5 | • | TC96263 | NP_187434 | 90.9/5.0 | 1.7 | 1 | 3/4 | M | |||
| 9 | HSP83 | −232 | • | • | TC106622 | 80.1/4.9 | 40 | 28 | 5/5 | C | |||
| 10 | HSP70 (cytosol) | −187 | • | • | • | TC100383 | 71.0/5.1 | 42 | 22 | 7/7 | C | ||
| −163 | • | • | • | TC100389 | 70.9/5.0 | 10 | 20 | 7/7 | C | ||||
| −240 | • | • | • | TC108198 | 71.5/5.2 | 50 | 27 | 7/7 | C | ||||
| −49 | • | • | BQ139103 | AAS57912 | 71.0/4.9 | 5.3 | 6 | 7/7 | C | ||||
| 11 | HSP70 (endoplasmic reticulum) | −63 | • | • | • | TC100382 | 73.5/5.0 | 12 | 9 | 4/5 | ER | ||
| −7 | • | TC100386 | CAB72128 | 73.4/4.8 | 2.4 | 2 | 3/3 | ER | |||||
| 12 | HSP70 (chloroplasts) | −280 | • | • | • | TC108910 | CAA49147 | 75.5/5.0 | 31 | 30 | 2/2 | CP | |
| 13 | HSP70 (mitochondria) | −75 | • | TC106412 | 70.9/5.3 | 15 | 10 | 3/4 | CP | ||||
| −29 | • | TC106356 | 72.3/5.8 | 6.9 | 4 | 3/3 | M | ||||||
| 14 | Leukotriene-A4 hydrolase like | −49 | • | TC95368 | 68.6/5.0 | 12 | 7 | 3/7 | C | ||||
| 15 | CPN60-2 | −8 | • | • | TC100626 | CAA50217 | 61.0/5.4 | 3.7 | 2 | 6/6 | M | ||
| −9 | • | • | TC100627 | CAA50217 | 61.0/5.4 | 3.1 | 2 | 6/6 | M | ||||
| 16 | Arg-methyltransferase | −10 | • | TC110222 | NP_850528 | 59.7/4.8 | 20 | 2 | 3/7 | C | |||
| −9 | • | TC95822 | NP_850528 | 59.7/4.8 | 4.3 | 2 | 3/7 | C | |||||
| 17 | TCP CPN60 chaperonin subunit ɛ§ | −3 | • | TC106794 | 52.9/5.3 | 3.8 | 1 | 7/9 | C | ||||
| 18 | TCP CPN60 chaperonin subunit τ | −32 | • | TC96153 | Q75HJ3 | 58.7/5.0 | 20 | 5 | 7/9 | C | |||
| 19 | ATPase subunit α (vacuole) | −9 | • | TC106363 | Q9SM09 | 68.6/5.1 | 4 | 2 | 5/5 | C | |||
| 20 | PDI | −311 | • | • | • | TC106707 | 57.0/4.9 | 54 | 34 | 4/4 | ER | ||
| 21 | Calreticulin | −63 | • | • | TC106673 | 48.4/4.4 | 15 | 8 | 3/3 | ER | |||
| 22 | Tubulin β | −133 | • | • | TC106434 | 50.5/4.7 | 38 | 14 | 8/10 | C | |||
| 23 | Tubulin α | −33 | • | TC100409 | 49.5/4.8 | 13 | 5 | 11/11 | C | ||||
| 24 | CPN60 subunit α | −345 | • | • | TC100730 | 61.9/4.8 | 61 | 46 | 1/1 | CP | |||
| 25 | CPN60 subunit β | −34 | • | TC100508 | 62.7/5.3 | 8 | 5 | 6/6 | CP | ||||
| 26 | ATP synthase subunit β (mitochondria) | −180 | •* | • | TC93945 | 59.8/5.6 | 37 | 19 | 2/2 | M | |||
| −5 | • | CX526674 | O04275 | 60.1/6.5 | 6.3 | 1 | 1/3 | M | |||||
| 27 | UDP-Glc pyrophosphorylase | −136 | • | TC94328 | 51.4/5.0 | 26 | 15 | 2/2 | C | ||||
| 28 | Pyruvate decarboxylase | −35 | • | TC100918 | 63.1/4.4 | 11 | 5 | 14/15 | C | ||||
| −116 | • | TC100561 | 65.0/5.6 | 29 | 13 | 14/17 | C | ||||||
| 29 | Phosphoglycerate dehydrogenase | −5 | • | NM_101636 | 66.4/5.6 | 1.3 | 2 | 3/6 | CP | ||||
| −15 | • | NM_119583 | 63.3/6.0 | 4.8 | 3 | 3/5 | CP | ||||||
| 30 | Ketol acid reductoisomerase§ | −3 | • | TC94305 | 62.9/6.2 | 2.2 | 1 | 6/7 | CP | ||||
| 31 | Succinate dehydrogenase | −15 | • | TC100922 | NM_126074 | 69.6/5.8 | 3.3 | 2 | 8/11 | M | |||
| 32 | Stress-induced protein (Sti1) | −40 | • | TC102206 | Q9STH1 | 63.7/5.8 | 13 | 6 | 6/6 | C | |||
| 33 | Selenium-binding protein | −90 | • | • | TC107269 | 54.2/5.5 | 28 | 11 | 7/7 | C | |||
| 34 | UDP-Glc dehydrogenase | −5 | • | TC106325 | 52.9/6.7 | 4.2 | 2 | 10/11 | C | ||||
| 35 | Succinic semialdehyde dehydrogenase (mitochondria) | −108 | • | • | TC101970 | 52.6/5.4 | 27 | 12 | 5/7 | CP | |||
| 36 | 4-Coumarate-CoA ligase | −21 | • | TC95226 | 55.8/5.5 | 7 | 4 | 4/6 | C | ||||
| 37 | Leu aminopeptidase | −100 | • | TC94615 | Q8GZD8 | 60.2/7.7 | 25 | 12 | 5/10 | CP | |||
| 38 | Vicilin (47 kD) | −180 | • | • | • | TC100301 | 53.1/5.1 | 29 | 21 | 1/2 | ER | ||
| −121 | • | • | • | TC100302 | 53.7/5.5 | 28 | 14 | 1/1 | ER | ||||
| 39 | PM51 | −6 | • | • | TC100258 | 55.1/5.7 | 3.4 | 2 | 1/1 | ER | |||
| 40 | SBP65 | −6 | • | TC102224 | Q41060 | 59.5/5.7 | 4.6 | 2 | 0/2 | C | |||
| 41 | Suc-binding protein P54 | −106 | • | • | • | TC94339 | 55.8/5.9 | 21 | 13 | 6/7 | ER | ||
| 42 | Malic enzyme (cytosol) | −136 | • | • | TC106706 | 65.3/5.9 | 33 | 16 | 5/6 | C | |||
| 43 | Aldehyde dehydrogenase | −97 | • | • | TC108200 | Q84V96 | 59.0/8.2 | 19 | 11 | 6/7 | M | ||
| −39 | • | • | TC106752 | 60.2/7.9 | 3.9 | 5 | 7/8 | M | |||||
| 44 | Rubisco large subunit | −46 | • | AJ847602 | O03042 | 52.9/5.7 | 18 | 5 | 9/9 | CP | |||
| 45 | RNA-binding protein | −65 | • | • | TC94137 | 39.4/6.7 | 17 | 7 | 1/1 | N | |||
| −25 | • | • | TC94494 | 40.5/6.1 | 6.3 | 4 | 1/1 | N | |||||
| 46 | Catalase | −15 | • | TC100988 | CAA42736 | 57.3/6.7 | 6.2 | 3 | 3/5 | C | |||
| 47 | EF1 subunit α | −145 | • | • | • | TC106485 | 49.1/9.0 | 8.2 | 16 | 6/6 | C | ||
| 48 | 26S proteasome regulatory subunit 6A | −89 | • | • | • | TC94498 | 47.4/4.7 | 24 | 10 | 4/4 | C | ||
| 49 | PDI like§ | −4 | • | TC106884 | 40.1/5.1 | 5.5 | 1 | 6/6 | ER | ||||
| 50 | Actin | −66 | • | • | • | • | TC106785 | 41.7/5.2 | 14 | 7 | 4/4 | C | |
| −94 | • | • | • | TC107326 | 41.7/5.3 | 23 | 11 | 4/4 | C | ||||
| −5 | • | TC109603 | XP_469569 | 41.8/5.1 | 5.7 | 2 | 4/4 | C | |||||
| −47 | • | • | • | TC106786 | XP_469569 | 41.8/5.1 | 2.7 | 7 | 4/4 | C | |||
| 51 | 4-Methyl-5(b-hydroxyethyl)- thiazole monophosphate biosynthesis protein | −185 | • | • | TC107759 | 46.8/6.7 | 38 | 20 | 3/3 | CP | |||
| 52 | SNF4b | −50 | • | • | AAO61675 | 41.9/5.3 | 31 | 7 | 5/6 | C | |||
| 53 | EF-tu | −133 | • | TC100566 | 53.1/6.1 | 26 | 14 | 1/2 | CP | ||||
| 54 | Gln synthetase (cytosol) | −61 | • | • | • | TC106729 | 39.1/5.2 | 24 | 7 | 3/3 | C | ||
| −56 | • | • | TC106808 | P04771 | 39.2/5.2 | 26 | 7 | 3/3 | C | ||||
| 55 | Gln synthetase (chloroplasts) | −33 | • | TC106913 | 47.0/6.2 | 16 | 5 | 3/6 | CP | ||||
| 56 | Legumin A | −96 | • | • | • | TC100259 | Q41702 | 56.5/6.5 | 7.9 | 12 | 6/7 | ER | |
| −137 | • | • | • | • | TC100252 | 60.4/6.2 | 23 | 16 | 5/7 | ER | |||
| 57 | Legumin J | −45 | • | •* | • | TC100250 | P05692 | 56.9/5.5 | 15 | 6 | 5/6 | ER | |
| −70 | • | •* | • | TC100253 | P05692 | 56.9/5.5 | 21 | 8 | 5/6 | ER | |||
| 58 | Enolase | −140 | • | TC100309 | 47.8/5.5 | 30 | 14 | 3/4 | C | ||||
| 59 | Phosphoglycerate mutase | −107 | • | TC100321 | 60.6/5.4 | 25 | 13 | 3/3 | C | ||||
| 60 | Allergen 28 K | −60 | • | • | • | TC101005 | 50.7/5.4 | 18 | 7 | 2/3 | C | ||
| 61 | Aldose-1-epimerase | −66 | • | TC95178 | 35.7/5.6 | 20 | 8 | 2/3 | C | ||||
| 62 | Cinnamoyl-CoA reductase | −4 | • | TC101159 | 36.0/5.7 | 5.3 | 2 | 7/8 | C | ||||
| 63 | 26S proteasome subunit 7 | −9 | • | TC109249 | O24412 | 34.7/6.0 | 15 | 2 | 0/1 | C | |||
| 64 | Chalcone reductase | −63 | • | TC100398 | 34.8/5.9 | 33 | 8 | 2/3 | C | ||||
| 65 | Glycinin subunit G7 | −31 | • | • | AJ498697 | Q6DR94 | 60.5/6.6 | 29 | 5 | 1/10 | ER | ||
| 66 | Aldo/keto auxin- induced reductase | −16 | • | TC94944 | 37.9/5.9 | 7.9 | 3 | 4/6 | C | ||||
| 67 | GAPDH (cytosol) | −349 | • | • | • | TC106518 | 36.6/6.5 | 53 | 34 | 2/2 | C | ||
| −134 | • | • | • | TC106519 | 37.0/6.9 | 26 | 14 | 2/2 | C | ||||
| 68 | Malate dehydrogenase | −78 | • | • | TC100430 | 35.6/6.1 | 25 | 9 | 5/5 | C | |||
| −78 | • | • | TC100429 | 35.6/6.1 | 27 | 10 | 5/6 | C | |||||
| 69 | Dehydrin | −160 | • | • | • | TC100921 | 31.2/6.4 | 41 | 14 | 0/1 | C | ||
| 70 | GTP-binding protein | −81 | • | • | TC95094 | Q9M7P3 | 44.4/6.3 | 40 | 9 | 5/7 | C | ||
| −44 | • | • | TC103575 | Q9M7P3 | 44.4/6.3 | 15 | 6 | 5/7 | C | ||||
| 71 | Ran | −113 | • | TC93955 | 25.2/6.3 | 51 | 12 | 6/6 | C | ||||
| 72 | 5,10-Methylene tetrahydrofolate dehydrogenase§ | −6 | • | TC107067 | 31.5/7.0 | 3.7 | 1 | 3/3 | C | ||||
| 73 | PM34 | −187 | • | • | • | • | TC107621 | 31.8/6.9 | 58 | 20 | 2/5 | C | |
| −57 | • | • | BQ122292 | Q9LLQ6 | 31.7/6.5 | 59 | 7 | 1/5 | C | ||||
| 74 | G-protein subunit β | −155 | • | • | • | TC100539 | 35.6/7.0 | 41 | 17 | 7/7 | C | ||
| 75 | ESP | −207 | • | • | TC103893 | 27.4/6.7 | 72 | 22 | 3/5 | C | |||
| 76 | GST (mitochondria) | −57 | • | TC95046 | 24.8/6.1 | 30 | 7 | 2/4 | M | ||||
| 77 | 1-Cys Prx | −196 | • | • | • | • | TC108877 | 24.3/6.0 | 57 | 22 | 2/3 | C | |
| 78 | EF1 subunit β | −129 | • | • | TC94335 | 24.2/4.4 | 39 | 15 | 1/1 | C | |||
| 79 | EF1 subunit δ | −86 | • | • | TC106814 | 25.3/4.2 | 43 | 11 | 1/1 | C | |||
| −48 | • | • | TC106815 | 25.3/4.2 | 4.4 | 7 | 1/1 | C | |||||
| 80 | DNA polymerase d auxiliary protein | −20 | • | AJ497831 | O82134 | 29.4/4.4 | 22 | 3 | 5/5 | N | |||
| 81 | PM24 | −43 | • | • | AW559621 | Q9SEL0 | 26.8/4.9 | 48 | 6 | 1/2 | N | ||
| 82 | Ran-binding protein | −33 | • | • | TC94495 | 24.5/4.5 | 16 | 5 | 4/4 | N | |||
| 83 | PM25 | −134 | • | • | • | • | ABB16353 | 26.4/4.5 | 69 | 15 | 1/1 | N | |
| 84 | 14-3-3 protein§ | −6 | • | TC106436 | 29.3/4.5 | 7.8 | 1 | 2/2 | C | ||||
| 85 | UK3 | −16 | • | TC108880 | 27.3/4.5 | 13 | 3 | 1/1 | M | ||||
| 86 | LEA protein (group 5) | −57 | • | TC96862 | P09444 | 26.9/4.8 | 40 | 7 | 0/1 | C | |||
| 87 | Wali 7 | −32 | • | TC100852 | 26.9/5.1 | 17 | 5 | 4/4 | C | ||||
| −45 | • | TC100620 | 28.0/5.6 | 17 | 6 | 4/6 | C | ||||||
| 88 | 2-Cys Prx | −122 | • | • | TC94151 | 29.1/5.8 | 34 | 14 | 2/3 | CP | |||
| −82 | • | • | TC93962 | 29.0/6.0 | 11 | 10 | 2/4 | CP | |||||
| 89 | Ferritin | −84 | • | • | TC96963 | Q1SCB6 | 20.3/5.9 | 30 | 10 | 3/4 | CP | ||
| 90 | UK4 | −20 | • | • | TC109465 | – | 22 | 3 | |||||
| 91 | Isopropylmalate dehydratase | −9 | • | TC101142 | 26.4/5.6 | 6.8 | 2 | 3/4 | CP | ||||
| 92 | SOUL-heme-binding protein | −76 | • | TC107473 | 25.3/5.4 | 20 | 9 | 2/3 | ER | ||||
| 93 | Triose phosphate isomerase | −43 | • | TC93925 | Q6GW08 | 27.2/5.6 | 20 | 6 | 3/4 | C | |||
| 94 | Sorbitol-6-P dehydrogenase§ | −4 | • | TC101758 | AAL86677 | 34.9/6.7 | 4.5 | 1 | 6/6 | C | |||
| 95 | Ni-binding urease accessory protein§ | −3 | • | TC94960 | 29.9/5.9 | 6.9 | 1 | 3/3 | C | ||||
| 96 | Short-chain alcohol dehydrogenase | −11 | • | TC106262 | Q9SQF9 | 28.1/5.1 | 13 | 2 | 4/5 | C | |||
| 97 | Gly-rich protein | −90 | • | • | TC98399 | Q75QN8 | 21.5/5.5 | 59 | 10 | 6/9 | C | ||
| 98 | Prx IIE | −7 | • | • | TC95578 | 23.2/9.6 | 8.9 | 2 | 2/2 | CP | |||
| 99 | Gly-rich RNA-binding protein | −32 | • | TC96436 | P49311 | 14.8/4.7 | 33 | 4 | 1/1 | C | |||
| −14 | • | TC100232 | P49311 | 16.3/4.9 | 21 | 3 | 1/1 | C | |||||
| −34 | • | • | TC93939 | Q41518 | 15.7/5.0 | 2.3 | 5 | 1/1 | C | ||||
| 100 | PM28 | −18 | • | • | CA989687 | Q9LE44 | 9.5/5.5 | 14 | 3 | 0/1 | C | ||
| 101 | Small HSP (18.2 kD) | −16 | • | TC100459 | 18.0/5.0 | 23 | 3 | 0/1 | C | ||||
| −16 | • | TC108710 | 17.1/4.9 | 17 | 3 | 0/1 | C | ||||||
| 102 | 2S albumin | −23 | • | • | • | TC100254 | 16.8/6.2 | 6.5 | 3 | 8/8 | ER | ||
| 103 | EIF5 A-2 | −95 | • | • | TC94209 | 17.2/5.1 | 70 | 11 | 4/4 | C | |||
| −51 | • | TC94697 | 17.5/5.2 | 14 | 7 | 4/4 | C | ||||||
| 104 | GSH peroxidase | −72 | • | • | TC94412 | 25.8/9.8 | 23 | 10 | 3/4 | M | |||
| 105 | Prx IIF§ | −5 | • | TC107761 | 21.4/9.4 | 4.9 | 1 | 2/2 | M | ||||
| 106 | Prx II | −81 | • | • | • | TC106854 | 17.5/5.5 | 40 | 9 | 2/2 | C | ||
| 107 | Ripening-related protein | −50 | • | TC106511 | 18.2/6.0 | 24 | 6 | 2/4 | C | ||||
| 108 | Nucleoside diphosphate kinase | −47 | • | TC106697 | 16.4/6.2 | 23 | 6 | 0/1 | C | ||||
| 109 | UK 5 | −62 | • | TC106727 | 22.6/9.7 | 27 | 8 | 1/1 | C | ||||
| 110 | 40S ribosomal protein S21 | −14 | • | TC98256 | 9.02/8.0 | 14 | 3 | 1/2 | C | ||||
| 111 | FKBP | −159 | • | TC106311 | 18.2/8.3 | 49 | 16 | 4/4 | C | ||||
| 112 | Nonspecific LTP | −9 | • | TC94138 | 11.3/9.0 | 14 | 2 | 9/10 | ER | ||||
| 113 | Conglutin | −123 | •* | AJ498597 | CAA46552 | 48.9/9.0 | 46 | 13 | 9/12 | ER | |||
| 114 | Sali 3.2 | −41 | •* | TC106693 | 30.5/6.0 | 20 | 6 | 6/6 | ER | ||||