TABLE 1.
List of Toxoplasma proteins putatively enriched at the parasitophorous vacuole membranea
| Gene ID (TGGT1_) | Description | mT-RAH + biotin/mT-RAH-biotin log2 FC | Evidence of PV/PVM/exported localization within infected cells | Predicted subcellular localization within tachyzoites (hyperLOPIT)b |
|---|---|---|---|---|
| 269950 | Hypothetical protein (GRA61) | 6.73 | PV/PVM (this study) | Dense granules |
| 220240 | GRA31 | 6.23 | PV/PVM (16) | Dense granules |
| 231210 | Sarcalumenin/eps15 family protein | 5.06 | NA | Nucleus-chromatin |
| 279350 | Hypothetical protein | 4.73 | NA | Dense granules |
| 215360 | Hypothetical protein (GRA62) | 4.48 | PV/PVM (this study) | Dense granules |
| 310780 | GRA4 | 4.44 | PV/PVM (30) | Dense granules |
| 216180 | Hypothetical protein | 4.42 | No (this study) | ER |
| 309590 | ROP1 | 4.40 | PV/PVM (62) | Rhoptries 1 |
| 217530 | Hypothetical protein (GRA63) | 4.21 | PV/PVM (this study) | Dense granules |
| 410370 | MAF1-related | 4.17 | NA | NA |
| 258580 | ROP17 | 4.17 | PV/PVM (12) | Rhoptries 1 |
| 210370 | ROP54 | 4.13 | PV/PVM (13) | Rhoptries 1 |
| 207920 | Hypothetical protein | 4.11 | NA | Unknown |
| 229010 | RON4 | 4.10 | PV/PVM (51) | Rhoptries 1 |
| 306030 | Glutathione s-transferase, N-terminal domain-containing protein | 4.07 | NA | ER |
| 203600 | GRA50 | 4.07 | PV/PVM (32) | Dense granules |
| 249530 | Putative exportin 1 | 4.06 | NA | Cytosol |
| 289530 | Ribosomal protein RPL19 | 4.03 | NA | 60S ribosome |
| 411470 | MAF1-related | 4.02 | NA | NA |
| 292130 | Ribosomal protein RPL13A | 4.02 | NA | 60S ribosome |
| 215470 | Ribosomal protein RPL10A | 3.99 | NA | 60S ribosome |
| 230980 | Myosin I | 3.97 | NA | Unknown |
| 311470 | RON5 | 3.94 | NA | Rhoptries 1 |
| 300100 | RON2 | 3.91 | NA | Rhoptries 1 |
| 286450 | GRA5 | 3.68 | PV/PVM (63) | Dense granules |
| 270240 | MAG1 | 3.67 | PV/PVM (64) | Dense granules |
| 254720 | GRA8 | 3.64 | PV/PVM (31) | Dense granules |
| 258870B | Hypothetical protein | 3.64 | NA | NA |
| 411430 | ROP5 | 3.62 | PV/PVM (29) | NA |
| 306338B | Putative dynein gamma chain, flagellar outer arm | 3.58 | NA | NA |
| 306060 | RON8 | 3.56 | Nascent PVM (50) | Rhoptries 1 |
| 208830 | GRA16 | 3.42 | PV/exported (65) | Dense granules |
| 220950 | MAF1b | 3.40 | PV/PVM (27) | Dense granules |
| 314810 | Ribosomal protein RPL7 | 3.31 | NA | 60S ribosome |
| 226430 | Reticulon protein | 3.19 | NA | ER |
| 309120 | Ribosomal protein RPL4 | 3.05 | NA | 60S ribosome |
| 215220 | GRA22 | 3.01 | PV/PVM (66) | Dense granules |
| 254470 | MYR1 | 3.00 | PV/PVM (67) | Dense granules |
| 291930 | RNA recognition motif-containing protein | 2.96 | NA | Cytosol |
| 288650 | GRA12 | 2.96 | PV/PVM (68) | Dense granules |
The relative abundance of each protein was determined across all samples using the peptide-level TMT quantification, and the log2 fold change (log2 FC) was determined for various sample ratios. The proteins were filtered by a minimum log2 FC of 2.95 for the (mT-RAH + Toxo)/(mT-RAH + Toxo no biotin) ratio and a log2 FC of −0.65 for the (mT-RAH + Toxo)/(mT + Toxo) ratio. Displayed here are all the Toxoplasma proteins (identified by at least 2 unique peptides, with a protein score of >25, and with protein coverage of >1%) which passed the above thresholds, ranked according to the log2 FC between the averaged (mT-RAH + Toxo)/(mT-RAH + Toxo no biotin) samples, the majority Toxoplasma identifiers (TGGT1_; i.e., the proteins that contain at least half of the peptides belonging to a group of proteins that cannot be unambiguously identified by unique peptides), and the descriptive name for each protein (“Description” column). If the protein had been previously localized to the PV/PVM or exported (via microscopy), the reference is indicated. NA, No microscopy evidence of PV/PVM localization or no predicted subcellular localization within tachyzoites.
The predicted subcellular localization of these proteins within tachyzoites based on a recent hyperLOPIT proteomics study is included for reference (25). The hypothetical proteins of unknown location and function that were chosen for follow-up localization studies are bolded.