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Immunoprecipitation studies confirmed that the wild-type WNK4 robustly bound to KLHL3, but interaction with either disease-causing WNK4 mutant tested was strikingly impaired.
Focusing on DKK1, one of the most robustly bound RBM47 target transcripts identified by HITS-CLIP, we investigated the possible role of RBM47 as a modulator of mRNA abundance.
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Using yeast two hybrid methodology, we have found that alpha arrestin tails robustly bind the WW domains of HECT E3 ligases and of other proteins (F.-C. Hsieh, W.-K. Chen and C. A., unpublished).
Consistent with the reported finding, an elongated α-helix-rich region of AMOT (residues 404-728 of AMOT-p130, Figure 4A) robustly binds to the entire C-terminal half of Merlin, forming a stable complex with 1 1 stoichiometry.
Daple bound robustly to FZD7R, and only weakly to others, indicating that Daple may engage preferentially with FZD7R.
We found that Slit bound robustly to cultured Drosophila cells expressing transgenic full-length Robo1, but interacted only weakly with untransfected cells or cells expressing Robo1ΔIg1.
Consistent with this, we were unable to detect binding between RanGTP and either MtNup188 or MtNup192 by gel filtration analysis whereas the NTR ScKap95 bound robustly to RanGTP.
Recombinant purified GST-Gαi3 bound robustly to purified His-Daple CT (aa 1650 2028, containing the GBA motif) when loaded with GDP (inactive) but not when loaded with GDP/AlF4− or GTPγS (both mimic the GTP-bound active G protein).
The strategy resulted in subnanomolar inhibitors that bind robustly across a clinically derived panel of drug-resistant variants.
At therapeutically relevant concentrations, Pt II) also binds robustly within conserved cation-binding pockets in Domains V and VI rRNA at the peptidyl transferase center.
Both the wildtype FOXL2 and FOXL2 C134W proteins bound the wildtype DNA probe robustly and bound the mutated probe at a reduced level (Fig. 5A and Fig. S2).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com