The Calpain-7 protease functions together with the ESCRT-III protein IST1 within the midbody to regulate the timing and completion of abscission.
Paine EL, Skalicky JJ, Whitby FG, Mackay DR, Ullman KS, Hill CP, Sundquist WI
Elife. 2023 Sep 29;12:e84515. doi: 10.7554/eLife.84515.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 193031 | pCA528 CAPN7 (1-75) | Bacterial expression plasmid for CAPN7 (1-75) N-terminal MIT domain |
| 193032 | pCA528 CAPN7 (81-165) | Bacterial expression plasmid for CAPN7 (81-165) C-terminal MIT domain |
| 193033 | pCA528 CAPN7 L61D (1-165) | Bacterial expression plasmid for CAPN7 (1-165) tandem MIT domains with L61D mutation |
| 193034 | pCA528 CAPN7 V18D,F98D (1-165) | Bacterial expression plasmid for CAPN7 (1-165) tandem MIT domains with V18D and F98D mutations that abolish binding to IST1. |
| 193035 | pCA528 IST1 L328D (316-366)(C-Cys) | Bacterial expression plasmid for IST1 (316-366) with C-terminal Cysteine for fluorescence polarization binding assays. Contains L328D mutation that diminishes binding to MIT domains. |
| 193036 | pCA528 IST1 L355A (316-366)(C-Cys) | Bacterial expression plasmid for IST1 (316-366) with C-terminal Cysteine for fluorescence polarization binding assays. Contains L355A mutation that diminishes binding to MIT domains. |
| 193037 | pCA528 IST1 L328D,L355A (316-366)(C-Cys) | Bacterial expression plasmid for IST1 (316-366) with C-terminal Cysteine for fluorescence polarization binding assays. Contains L328D and L355A mutations that diminishes binding to MIT domains. |
| 193038 | pCA528 IST1 (322-366) | Bacterial expression plasmid for IST1 (322-366), the minimal construct with full binding to CAPN7 MIT domains. |
| 193039 | pCA528 IST1 (1-366) | Bacterial expression plasmid for full-length IST1 (1-366). |
| 193040 | pCA528 IST1 L328D, L355A (1-366) | Bacterial expression plasmid for full-length IST1 (1-366) with L328D and L355A mutations that diminish binding to MIT domains. |
| 193041 | pCAG-CAPN7-OSF | Mammalian expression plasmid encoding full-length CAPN7 with C-terminal OSF tag (One-Strep-FLAG) |
| 193042 | pCAG CAPN7 V18D-OSF | Mammalian expression plasmid encoding full-length CAPN7 with V18D mutation that diminishes binding to IST1. Contains C-terminal OSF tag (One-Strep-FLAG) |
| 193043 | pCAG CAPN7 L61D-OSF | Mammalian expression plasmid encoding full-length CAPN7 with L61D mutation. Contains C-terminal OSF tag (One-Strep-FLAG) |
| 193044 | pCAG CAPN7 F98D-OSF | Mammalian expression plasmid encoding full-length CAPN7 with F98D mutation that diminishes binding to IST1. Contains C-terminal OSF tag (One-Strep-FLAG) |
| 193045 | pCAG CAPN7 V18D,F98D-OSF | Mammalian expression plasmid encoding full-length CAPN7 with V18D and F98D mutations that diminishes binding to IST1. Contains C-terminal OSF tag (One-Strep-FLAG) |
| 193046 | pLVX-CAPN7-V18D-mCherry | Lentiviral expression vector for stable integration of inducible, siRNA-resistant CAPN7 with V18D (IST1-binding mutation) with C-terminal mCherry tag. |
| 193047 | pLVX-CAPN7-C290S-mCherry | Lentiviral expression vector for stable integration of inducible, siRNA-resistant CAPN7 with C290S (proteolysis inactivating mutation) with C-terminal mCherry tag. |