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The MAP4(Ala) mutant, which mimicked the dephosphorylated form, suppressed mitochondrial translocation and apoptosis.
Nevertheless, it was observed that the Aqp1b-S254A mutant prevented phosphorylation and increased Aqp1b translocation into the plasma membrane and subsequent water permeability, whereas the Aqp1b-S254D mutant, which mimicked the constitutively phosphorylated state of Aqp1b, was predominantly located in intracellular vesicles.
On the other hand, the UCP3RE171/172GG mutant, which mimicked the wild-type UCP3 exclusively for mitochondrial uptake of entering Ca2+ in intact cells, exhibited its activity in digitonin-permeabilized cells only at low Ca2+ concentrations while it appeared to be inactive at high Ca2+ concentrations.
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To this end, we designed the recombinant Apaf-1ΔWD40 S268D mutant, which mimics Ser268 phosphorylation.
Binding assays, using the Escherichia coli FabF C163Q mutant, which mimics the acyl-enzyme intermediate9, revealed that the dissociation constants (KD) for 1 and 2 were ~2-fold tighter than those of 3 and 4 (Table 2 and Supplementary Figs. 38b and 39).
Staining of the triple mutant which mimics the binding site of the HLA-C2 allotypes (i.e. 221/HLA-G M76V+Q79R+T80K) resulted in a strong binding of the KIR2DL1-Ig (Fig. 3, bold black frame).
The triple HLA-G mutant which mimics the binding site of the HLA-C1 allotypes (i.e. 221/HLA-G M76V+Q79R+T80N) was strongly stained with KIR2DL2-Ig (Fig. 3, bold black frame) but not with KIR2DL1-Ig.
However, when the 221/HLA-G mutants are assayed with KIR2DL1+ NK clones, only the triple mutant which mimics the binding site of the HLA-C2 allotypes (i.e. 221/HLA-G M76V+Q79R+T80K) inhibits the killing, in a similar manner to that observed with 221/HLA-Cw4 221/HLA-Cw4-Cw6 (Fig. 4B, dand bars).
When the various HLA-G mutants were incubated with KIR2DL2+ NK clones only the triple mutant which mimics the binding site of the HLA-C1 allotypes (i.e. 221/HLA-G M76V+Q79R+T80N) inhibits the killing similar to that of 221/HLA-Cw3 221/HLA-Cw3ark bars).
Transient transfection studies indicated that an EBP1 T261E mutant, which mimics EPB1 phosphorylated by PAK1, increased ErbB2 protein levels.
Transfection of an EBP1 mutant, which mimics PAK1-induced phosphorylation at T261 induces tamoxifen resistance in MCF-7 cells.
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