Table 1.
List of mouse ATGL variants designed for cloning and expression screening at the OPPF
| Fusion | N/C-term amino acid (POI) | Vector (pOPIN) | Design | Molecular weight (kDa) | Host tested | Host expressed |
|---|---|---|---|---|---|---|
| M1-Q257 | pOPINFS | HIS6-3C-POI-StrepII | 31 | B, I, M | — | |
| M1-E272 | pOPINFS | HIS6-3C-POI-StrepII | 33 | B, I, M | — | |
| M1-D288 | pOPINFS | HIS6-3C-POI-StrepII | 35 | B, I, M | — | |
| M1-K295 | pOPINFS | HIS6-3C-POI-StrepII | 36 | B | — | |
| M1-D364 | pOPINFS | HIS6-3C-POI-StrepII | 43 | B | — | |
| M1-H390 | pOPINFS | HIS6-3C-POI-StrepII | 46 | B | — | |
| M1-C486 | pOPINFS | HIS6-3C-POI-StrepII | 56 | B, I, M | — | |
| GST | M1-Q257 | pOPINJ | HIS6-GST-3C-POI | 56 | B, I, M | (B), I |
| GST | M1-E272 | pOPINJ | HIS6-GST-3C-POI | 58 | B, I | I |
| GST | M1-D288 | pOPINJ | HIS6-GST-3C-POI | 60 | B, I | (B), I |
| GST | M1-K295 | pOPINJ | HIS6-GST-3C-POI | 60 | B | (B) |
| GST | M1-D364 | pOPINJ | HIS6-GST-3C-POI | 68 | B | — |
| GST | M1-H390 | pOPINJ | HIS6-GST-3C-POI | 71 | B | — |
| GST | M1-C486 | pOPINJ | HIS6-GST-3C-POI | 81 | B, I, M | — |
| MBP | M1-Q257 | pOPINM | HIS6-MBP-3C-POI | 71 | B, I | B, I, (M) |
| MBP | M1-E272 | pOPINM | HIS6-MBP-3C-POI | 73 | B, I | B, I |
| MBP | M1-D288 | pOPINM | HIS6-MBP-3C-POI | 74 | B, I | B, I |
| MBP | M1-K295 | pOPINM | HIS6-MBP-3C-POI | 75 | B | B |
| MBP | M1-D364 | pOPINM | HIS6-MBP-3C-POI | 83 | B | (B) |
| MBP | M1-H390 | pOPINM | HIS6-MBP-3C-POI | 86 | B | (B) |
| MBP | M1-C486 | pOPINM | HIS6-MBP-3C-POI | 96 | B, I, M | I |
| NUSA | M1-Q257 | pOPINNUSA | HIS6-NUSA-3C-POI | 85 | B, I | B, I |
| NUSA | M1-E272 | pOPINNUSA | HIS6-NUSA-3C-POI | 87 | B, I | B, I |
| NUSA | M1-D288 | pOPINNUSA | HIS6-NUSA-3C-POI | 89 | B, I | B, I |
| NUSA | M1-K295 | pOPINNUSA | HIS6-NUSA-3C-POI | 89 | B | B |
| NUSA | M1-D364 | pOPINNUSA | HIS6-NUSA-3C-POI | 97 | B | (B) |
| NUSA | M1-H390 | pOPINNUSA | HIS6-NUSA-3C-POI | 100 | B | (B) |
| NUSA | M1-C486 | pOPINNUSA | HIS6-NUSA-3C-POI | 110 | B, I, M | I |
| SMT3 | M1-Q257 | pOPINS3C | HIS6-SMT3-3C-POI | 41 | B, I, M | (B), I, M |
| SMT3 | M1-E272 | pOPINS3C | HIS6-SMT3-3C-POI | 43 | B, I, M | (B), I, M |
| SMT3 | M1-D288 | pOPINS3C | HIS6-SMT3-3C-POI | 45 | B, I, M | B, I, M |
| SMT3 | M1-K295 | pOPINS3C | HIS6-SMT3-3C-POI | 46 | B | (B) |
| SMT3 | M1-D364 | pOPINS3C | HIS6-SMT3-3C-POI | 53 | B | (B) |
| SMT3 | M1-H390 | pOPINS3C | HIS6-SMT3-3C-POI | 56 | B | (B) |
| SMT3 | M1-C486 | pOPINS3C | HIS6-SMT3-3C-POI | 67 | B, I, M | (M) |
| TRX | M1-Q257 | pOPINTRX | HIS6-TRX-3C-POI | 42 | B, I, M | (B), (M) |
| TRX | M1-E272 | pOPINTRX | HIS6-TRX-3C-POI | 44 | B, I, M | (B) |
| TRX | M1-D288 | pOPINTRX | HIS6-TRX-3C-POI | 45 | B, I | (B), I |
| TRX | M1-K295 | pOPINTRX | HIS6-TRX-3C-POI | 46 | B | (B) |
| TRX | M1-D364 | pOPINTRX | HIS6-TRX-3C-POI | 54 | B | (B) |
| TRX | M1-H390 | pOPINTRX | HIS6-TRX-3C-POI | 57 | B | (B) |
| TRX | M1-C486 | pOPINTRX | HIS6-TRX-3C-POI | 67 | B, I, M | I |
Variants were tested for overexpression in E.coli “B,” the insect cell line Sf9 “I,” and the mammalian HEK293T cells “M.” In the last column, detectable, yet insoluble target protein of the protein is indicted in brackets, for example, (B).
Abbreviations: N/C-term amino acid, first and last amino acid of mATGL in the variant (not including any tags); POI, protein of interest.