TABLE 1.
Protein spots that were differentially expressed in epimastigote stage of TcI isolates of T. cruzia
| Protein spot | Protein name (gene) | NCBI accession no. | Mol mass (Da) | Peptide count | Protein score | Protein score (CI%) | pI | E value |
|---|---|---|---|---|---|---|---|---|
| Spots conserved in the three isolates | ||||||||
| 1 | Chain A, tyrosine aminotransferase (TcTAT) | gi|6137418 | 46,120.3 | 12 | 188 | 100 | 5.72 | 3.155E–13 |
| 3 | Tyrosine aminotransferase (TcTAT) | gi|71407311 | 46,238.5 | 13 | 442 | 100 | 5.82 | 1.256E–38 |
| 4 | Tyrosine aminotransferase N-term (TcTAT) | gi|71655339 | 27,698.9 | 12 | 400 | 100 | 5.71 | 1.991E–34 |
| 5 | l-Tyrosine:2-oxoglutarate aminotransferase (TcTAT) | gi|1168606 | 46,137.4 | 20 | 737 | 100 | 5.82 | 3.972E–68 |
| 6 | Tyrosine aminotransferase (TcTAT) | gi|71413938 | 46,100.4 | 16 | 537 | 100 | 5.66 | 3.972E–48 |
| 7 | NADPH flavin oxidoreductase (old yellow enzyme, TcOYE) | gi|61741942 | 42,191.6 | 19 | 902 | 100 | 6.03 | 1.256E–84 |
| 8 | NADPH flavin oxidoreductase (old yellow enzyme, TcOYE) | gi|61741950 | 42,233.6 | 18 | 889 | 100 | 5.97 | 2.506E–83 |
| 9 | l-Threonine 3-dehydrogenase (TcTDH) | gi|71406160 | 36,880.7 | 8 | 181 | 100 | 6.71 | 1.581E–12 |
| 10 | NADPH flavin oxidoreductase (old yellow enzyme, TcOYE) | gi|61741952 | 42,080.5 | 20 | 749 | 100 | 6.03 | 2.506E–69 |
| 11 | Aldo-keto reductase/quinone oxido-reductase (AKR) | gi|189396135 | 32,485.5 | 13 | 499 | 100 | 7.18 | 1.256E–43 |
| 12 | Aldo-keto reductase/quinone oxido-reductase (AKR) | gi|14279174 | 31,764.2 | 13 | 376 | 100 | 8.58 | 2.506E–31 |
| Protein spots differentially expressed in COL (compared to TCC) | ||||||||
| 13 | Proline racemase B (TcPRACB) | gi|74812220 | 38,440.6 | 12 | 442 | 100 | 5.44 | 1.256E–38 |
| 14 | Proline racemase N-term (TcPRACB) | gi|71400794 | 32,411.5 | 11 | 435 | 100 | 5.63 | 6.295E–38 |
| 15 | Major paraflagellar rod protein 2 (TcPAR2) | gi|162179 | 69,504.9 | 19 | 355 | 100 | 5.85 | 6.295E–30 |
| 16 | Paraflagellar rod protein N-term (TcPAR) | gi|71411387 | 42,471.9 | 16 | 355 | 100 | 6.12 | 6.295E–30 |
| 17 | Paraxonemal (paraflagellar) rod protein (TcPAR2) | gi|71398801 | 31,336.1 | 10 | 164 | 100 | 5.95 | 7.924E–11 |
| 18 | 2-Hydroxyacid dehydrogenase N-term, d-isomer (2-Hacid-dh) | gi|71406187 | 28,516.7 | 16 | 567 | 100 | 6.17 | 3.972E–51 |
| 20 | Short interspersed repetitive element (TcSIRE) | gi|1911172 | 11,565 | 10 | 530 | 100 | 5.08 | 1.991E–47 |
| 21 | 2-Hydroxyacid dehydrogenase, d-isomer (2-Hacid-dh) | gi|71406189 | 22,228.2 | 11 | 296 | 100 | 5.87 | 5E–24 |
| 23 | HpcH/HpaI aldolase (TcP28) | gi|4558456 | 29,886.2 | 7 | 156 | 100 | 6.06 | 5E–10 |
| 24 | Pyridoxal kinase (TcPDXK) | gi|71418579 | 33,273.6 | 4 | 125 | 100 | 5.93 | 6.295E–07 |
| 25 | NADP-dependent malic enzyme N term (TcME1) | gi|71662506 | 62,705.5 | 12 | 307 | 100 | 6.55 | 3.972E–25 |
| 26 | NADP-dependent malic enzyme N term (TcME1) | gi|71651465 | 62,634.4 | 14 | 300 | 100 | 6.37 | 1.991E–24 |
| 27 | Cytochrome c oxidase subunit IV (TcCOXIV) | gi|71660419 | 38,924.4 | 16 | 351 | 100 | 5.72 | 1.581E–29 |
| 28 | Tryparedoxin peroxidase (TcTXNPx) | gi|17224953 | 22,246.1 | 11 | 413 | 100 | 5.96 | 9.976E–36 |
| 29 | Tryparedoxin peroxidase (TcTXNPx) | gi|71399514 | 22,375.1 | 11 | 411 | 100 | 6.32 | 1.581E–35 |
| 30 | Tryparedoxin peroxidase (TcTXNPx) | gi|4388655 | 25,545.3 | 22 | 167 | 100 | 7.56 | 2.506E–28 |
| 31 | Calpain-like cysteine peptidase (TcCAPN1) | gi|71411006 | 12,798.3 | 6 | 179 | 100 | 5.07 | 1.256E–11 |
| 32 | Beta tubulin of T. cyclops (TcTUBB) | gi|91983197 | 44,496.8 | 13 | 410 | 100 | 5.68 | 1.991E–35 |
| 33 | Beta tubulin of T. danilewskyi (TdTUBB) | gi|74229926 | 49,695.8 | 12 | 391 | 100 | 4.74 | 1.581E–33 |
| 34 | Alpha tubulin of T. cyclops (TcTUBA) | gi|91983196 | 49,777.5 | 19 | 677 | 100 | 4.93 | 1.991E–61 |
| 35 | Alpha tubulin of T. brucei (TbTUBA) | gi|115504283 | 49,755.5 | 21 | 646 | 100 | 4.93 | 2.506E–58 |
| 36 | Elongation factor 1α (TcEF1A) | gi|71408910 | 49,083.4 | 19 | 416 | 100 | 8.93 | 5E–36 |
| 37 | Ribokinase (TcRBKS) | gi|71659743 | 35,262.6 | 7 | 158 | 100 | 6.95 | 3.155E–10 |
| 38 | Heat shock protein 70 partial (TcHSP70) | gi|10626 | 74,233.7 | 31 | 626 | 100 | 5.64 | 2.506E–56 |
| 39 | Heat shock protein HSP70 (TcHSP70) | gi|162117 | 71,210.1 | 30 | 623 | 100 | 5.42 | 5E–56 |
| 40 | Heat shock 70-kDa protein, mitochondrial (TcHSP70-10) | gi|71407515 | 70,946 | 24 | 516 | 100 | 5.75 | 2.506E–45 |
| Spots differentially expressed in SYL (compared to TCC) | ||||||||
| 53 | Cytochrome c oxidase subunit IV (TcCOXIV) | gi|71667854 | 38,884.4 | 15 | 339 | 100 | 5.72 | 2.506E–28 |
| 54 | Tryparedoxin peroxidase (TcTXNPx) | gi|71413207 | 22,376.1 | 10 | 357 | 100 | 5.69 | 3.972E–30 |
| 55 | Tryparedoxin peroxidase (TcTXNPx) | gi|4388655 | 25,506.1 | 6 | 134 | 100 | 7.62 | 7.924E–08 |
| 56 | Elongation factor alpha G5 (TcEFA-G5) | gi|52424046 | 49,067.4 | 18 | 405 | 100 | 8.93 | 6.295E–35 |
| 57 | Heat shock protein 70 (TcHSP70) | gi|50659756 | 71,329.9 | 30 | 524 | 100 | 5.06 | 3.972E–46 |
| 58 | Glucose-regulated protein 78 (TcHSPA5 or TcGRP78) | gi|71415505 | 71,271.9 | 29 | 510 | 100 | 5.09 | 9.976E–45 |
| 59 | Glucose-regulated protein 78 (TcHSPA5 or TcGRP78) | gi|349838 | 71,488 | 25 | 365 | 100 | 5.07 | 3.155E–30 |
| 75- to 77-kDa antigen (TcHSP70) | gi|1097750 | 41,716.5 | 17 | 273 | 100 | 4.99 | 5E–21 | |
| 60 | Mitochondrial heat shock protein (TcHSP70mt) | gi|205278870 | 70,969.1 | 25 | 527 | 100 | 5.87 | 1.99E–46 |
| 61 | Heat shock protein 70 (TcHSP70) | gi|205278868 | 71,193 | 26 | 517 | 100 | 5.32 | 2.51E–73 |
| 62 | Alpha tubulin (TcTUBA) | gi|71397525 | 49,767.5 | 22 | 806 | 100 | 4.94 | 2.51E–74 |
| 62 | Alpha tubulin (TcTUBA) | gi|1314208 | 46,927.4 | 20 | 787 | 100 | 5.49 | 1.99E–72 |
| 63 | Beta tubulin 1.9 (TcTUBB) | gi|18568139 | 49,667.8 | 16 | 643 | 100 | 4.74 | 5.00E–58 |
| 64 | Beta tubulin of T. cyclops (TcTUBB) | gi|91983197 | 44,496.8 | 14 | 626 | 100 | 5.68 | 2.51E–56 |
| 65 | Calpain-like cysteine peptidase (TcCAPN1) | gi|71409962 | 12,839.3 | 5 | 170 | 100 | 5.08 | 1.26E–11 |
| 66 | Tryparedoxin peroxidase family (TcGPX1) | gi|19171158 | 16,057.1 | 5 | 102 | 99.9 | 5.27 | 9.98E–51 |
| 67 | Histone H2B of T. brucei (TbHIST1H2B) | gi|71748436 | 12,588.8 | 4 | 84 | 96.1 | 11.77 | 1.26E–54 |
| 68 | Beta tubulin (TcTUBB) | gi|71656281 | 49,667.7 | 15 | 570 | 100 | 4.7 | 9.51E–56 |
| 69 | Alpha tubulin of T. danilewskyi (TdTUBA) | gi|74229924 | 49,765.6 | 21 | 722 | 100 | 4.94 | 6.29E–66 |
| 70 | Cofilin-actin depolymerizing factor (TcCFL1) | gi|71417693 | 15,735.8 | 9 | 300 | 100 | 5.69 | 9.98E–24 |
| 71 | Prostaglandin F synthase (TcPGFS) | gi|71651365 | 32,489.5 | 16 | 532 | 100 | 7.71 | 6.29E–47 |
| 72 | Hypothetical protein | gi|71657351 | 129,881.1 | 18 | 77 | 79.8 | 6.84 | 7.92E–41 |
| 73 | Lipophosphoglycan biosynthetic protein (TcLPG3) | gi|71663313 | 86,968.5 | 17 | 248 | 100 | 5.26 | 1.58E–18 |
| 74 | 3-Hydroxy-3-methylglutaryl coenzyme A synthase-like (TcHMGCS2) | gi|71661816 | 54,719.2 | 14 | 445 | 100 | 6.17 | 3.15E–38 |
| 75 | Carboxypeptidase (TcCPD) | gi|71667929 | 57,626.9 | 19 | 136 | 100 | 5.92 | 2.51E–07 |
| Metallocarboxypeptidase (TcCPD) | gi|47076074 | 57,686.9 | 18 | 127 | 100 | 5.85 | 1.99E–06 | |
a
T. cruzi epimastigotes (COL, SYL, and TCC isolates) were cultured in LIT media. Protein homogenates were resolved by two-dimensional gel electrophoresis. Gel images were analyzed on Progenesis SameSpots software, and normalized spot volumes were used to compare different groups. Proteins spots with a >2-fold change were subjected to MALDI-TOF MS/MS analysis. The number of peptides for each protein spot for which peptide mass fingerprint was obtained by MALDI-TOF MS is listed. Protein spots that were identified with high confidence by Mascot search engine (a protein score of >62) and statistically significant at the 95% confidence interval (CI%) are listed. The probability of match of the peptides in database search was accepted as significant at an E value of <0.001. pI, isoelectric pH. Underlined text indicates proteins involved in host cell invasion and cytoskeletal remodeling and motility. Boldface text indicates proteins of the antioxidant family involved in redox homeostasis.