TABLE 1.
Summary of the Duox and DuoxA expression constructs used
PoxH, peroxidase homology; F, first half; S, second half; N, N terminus; C, C terminus; A, A-loop; C, C-loop; E, E-loop; FLAG-tagged constructs are in p3×FLAG-CMV vector and all other constructs in pcDNA3.1.
| Name of protein | Structures of expressed protein |
|---|---|
| Duox | |
| Duox1 and Duox2 | Wild type Duox1 and Duox2 |
| 2×HA-Duox1 | 2 × HA tag between Asn-23 and Pro-24 of extracellular PoxH domain |
| 2×HA-Duox2 | 2 × HA tag between Asp-27 and Ala-28 of extracellular PoxH domain |
| Duox PoxH-domain chimera | |
| Duox(1PoxH-2) | Chimera of PoxH domain of Duox1 (1–592 aa) and Nox-like portion of Duox2 (596–1548 aa) |
| Duox(2PoxH-1) | Chimera of PH domain of Duox2 (1–594 aa) and Nox-like portion of Duox1 (592–1551 aa) |
| DuoxA | |
| DuoxA1 and DuoxA2 | Wild type DuoxA1 and DuoxA2 |
| 3NxFLAG-DuoxA1 | 3xFLAG tag at the extracellular N terminus |
| DuoxA1–3CxFLAG | 3xFLAG tag at the intracellular C terminus |
| DuoxA1–1CxFLAG | 1xFLAG tag at the extracellular C-terminus |
| DuoxA1(N-del) | The extracellular N terminus (2–17 aa) deletion |
| 3NxFLAG-DuoxA2 | 3xFLAG tag at the extracellular N terminus |
| 2NxFLAG-DuoxA2 | 2 × FLAG tag at the extracellular N terminus |
| 1NxFLAG-DuoxA2 | 1xFLAG tag at the extracellular N terminus |
| 2NxHA-DuoxA2 | 2 × HA tag at the extracellular N terminus |
| DuoxA2–3CxFLAG | 3xFLAG tag at the intracellular C terminus |
| DuoxA2(N-del) | The extracellular N terminus (2–17 aa) deletion |
| DuoxA N-terminal chimera | |
| DuoxA(1–2) | Chimera of extracellular N terminus of DuoxA1 (1–19 aa) and DuoxA2 (20–320 aa) |
| DuoxA(2–1) | Chimera of extracellular N terminus of DuoxA2 (1–19 aa) and DuoxA1 (20–343 aa) |
| DuoxA(1F-2) | Chimera of DuoxA1 (1–8 aa; first half of 19aa) and DuoxA2 (9–320 aa) |
| DuoxA(2–1S-2) | Chimera of DuoxA2 (1–11 aa), DuoxA1 (12–19 aa; second half of 19aa), and DuoxA2 (20–320 aa) |
| A-loop | |
| Duox2→1(A:5aa) | 5aa in A-loop of Duox2 is replaced by corresponding 5aa in A-loop of Duox1 |
| Duox2→1(A:8aa) | 8aa in A-loop of Duox2 is replaced by corresponding 8aa in A-loop of Duox1 |
| Duox1→2(A:F4aa) | 4aa (first half of 8aa) in A-loop of Duox1 is replaced by corresponding 4aa in A-loop of Duox2 |
| Duox1→2(A:S4aa) | 4aa (second half of 8aa) in A-loop of Duox1 is replaced by corresponding 4aa in A-loop of Duox2 |
| Duox1→2(A:8aa) | 8aa in A-loop of Duox1 replaced by corresponding 8aa in A-loop of Duox2 |
| Duox1→2(A:8aa+L) | 8aa + Arg in A-loop of Duox1 replaced by corresponding residues (8aa + Leu) in A-loop of Duox2 |
| Duox1→2(A:8aa+GL) | 8aa + Ala and Arg in A-loop of Duox1 replaced by corresponding residues (8aa + Gly and Leu) in A-loop of Duox2 |
| Duox1→2(A:8aa+L)+L/A | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Leu to Ala |
| Duox1→2(A:8aa+L)+PP/HH | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Pro-Pro to His-His |
| Duox1→2(A:8aa+L)+PP/HP | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Pro-Pro to His-Pro |
| Duox1→2(A:8aa+L)+PP/PH | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Pro-Pro to Pro-His |
| Duox1→2(A:8aa+L)+D/G | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Asp to Gly |
| Duox1→2(A:8aa+L)+A/T | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Ala to Thr |
| Duox1→2(A:8aa+L)+Q/D | Duox1→2 (A:8aa + L) with reversed mutation (Duox2 to Duox1), Gln to Asp |
| Duox1→2(A:PPD) | 3aa residues in A-loop of Duox1 are replaced by corresponding Pro,Pro and Asp residues in A-loop of Duox2 |
| Duox-Nox(A-loop) chimera | |
| Duox1→Nox1(A) | 7aa in A-loop of Duox1 replaced by corresponding residues in A-loop of Nox1 |
| Duox1→Nox5(A) | 7aa in A-loop of Duox1 replaced by corresponding residues in A-loop of Nox5 |
| Duox2→Nox1(A) | 7aa in A-loop of Duox2 replaced by corresponding residues in A-loop of Nox1 |
| Duox2→Nox5(A) | 7aa in A-loop of Duox2 replaced by corresponding residues in A-loop of Nox5 |
| C-loop | |
| Duox2→1(C:10aa) | 10aa (IVLAINVVNK) in C-loop of Duox2 replaced by corresponding 10 aa in C-loop of Duox1 (LIVSLGLHDE) |
| E-loop | |
| Duox2→1(E:TY/RF) | Thr and Tyr in E-loop of Duox2 replaced by corresponding Arg and Phe in E-loop of Duox1 |