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According to ITC analysis (Supplementary Table S1 and Figure S5), the six mutants bound to Cc with slightly larger KD values than 14-3-3ε FL WT.
These mutants bound to both FADD and caspase-8, but could not block apoptosis or the formation of death effector filaments.
We have determined the three-dimensional structures of selected mutants bound to the substrate analogue nicotinic acid, using X-ray crystallography.
All pyrin mutants bound to PSTPIP1 and reticularized PSTPIP1 filaments in a manner similar to wildtype pyrin (Figure 6A R).
With the exception of Et in complex with QacR(E120Q), wherein no drug density could be clearly ascribed and hence is not discussed further, electron density for each drug was clear and, as observed in previous wt QacR-drug complexes [3], [5], these mutants bound to each drug in a one drug per QacR dimer stoichiometry (data not shown).
All three mutants bound to PG like the wild-type, and no significant differences in the binding affinity were observed.
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In the presence of Ca2+, the wild-type protein and both mutants bind to both PS and phosphorylated PtdIns's with the exception of PtdIns 3,4,5 P3.
The profiles of proteins identified in wild-type and mutant Rab7 were qualitatively identical, suggesting that Rab7 mutants bind to the same complement of interactors as wild-type Rab7 (Fig. 4A and B, Supplementary Material, Fig. S4).
Instead, the newly termed "superbinder" Keap1 mutants bind to Nrf2 with a greater affinity than wild-type Keap1, yet are unable to inhibit Nrf2-mediated gene expression (Hast et al., 2014).
Here, we present the X-ray crystal structures of a dominant-negative form of human Scp1 (D96N mutant) bound to mono- and diphosphorylated peptides encompassing the CTD heptad repeat (Y1S2P3T4S5P6S7).
As shown in Figure 4D, Pc2 and the SIM1 deletion mutant bound to GST-SUMO1, whereas binding of the SIM2 or double SIM mutant was clearly reduced.
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