Protocol for deriving proximity, affinity, and stoichiometry of protein interactions using image-based quantitative two-hybrid FRET.
Feldmann C, Schanzler M, Ben-Johny M, Wahl-Schott C
STAR Protoc. 2023 Jul 28;4(3):102459. doi: 10.1016/j.xpro.2023.102459. PubMed Article
Feldmann C, Schanzler M, Ben-Johny M, Wahl-Schott C
STAR Protoc. 2023 Jul 28;4(3):102459. doi: 10.1016/j.xpro.2023.102459. PubMed Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 198192 | mVenus | Spectral correction: Expression of mVenus |
| 198196 | mTurquoise2 | Spectral correction: Expression of mTurquoise2 |
| 198197 | Dimer 3 AA | FRET calibration: Dimer construct consisting of mTurquoise2 and mVenus with a 3AA linker |
| 198198 | Dimer 42 AA | FRET calibration: Dimer construct consisting of mTurquoise2 and mVenus with a 42AA linker |
| 198199 | Dimer cleaved | FRET calibration: Dimer construct consisting of mTurquoise2 and mVenus with a 2A peptide linker for posttranslational linker cleavage |
| 198200 | mTq2-Calmodulin | FRET donor: Calmodulin-2 tagged with N-terminal mTurquoise2 |
| 198201 | mVenus-IQ6 WT | FRET acceptor: IQ6 domain of Moysin Va tagged with N-terminal mVenus |
| 198202 | mVenus-IQ6 R6A | FRET acceptor: Mutated IQ6 domain of Moysin Va tagged with N-terminal mVenus (lower binding affinity) |