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B56γ1 R201A mutation completely abolished the binding of the BubR1 pSer676-phosphopeptide (Table 2 and Fig. S5), likely because mutation of the semi-buried B56 Arg201 induced major local conformational changes, which in turn affected the interaction of the critical BubR1 Glu674 residue in positively charged pocket (Figs. 2 and S5).
Because mutation of the GATA element failed to reduce DNA binding in electromobility shift analyses (Fig. 4B), and because mutation of the Ebox reduced the AICAR effect to that of control values (Fig. 4C), we believe that PGC-1α expression is more likely to be regulated by USF-1 rather than GATA-4 in response to AICAR.
Furthermore, the high ATPase activity measured here for BmrC/BmrD is specific because mutation of the invariant lysine residue to an alanine in the Walker-A motifs of both BmrC and BmrD led to an affinity-purified mutant transporter with a low residual ATPase activity in detergent (cf. Fig. S2b).
This is because mutation of the ECM-binding sites has significantly perturbed the ability of PR/NMB to bind to all three ECM components.
The E3 ligase activity of TRAF6 is likely important for IRF3 activation because mutation of the RING domain in TRAF6 abolishes its activity.
Because mutation of the DES1 gene disrupts H2S generation in the Arabidopsis cytosol, restoring the capacity of H2S generation eliminates the phenotypic differences of the null mutants from wild-type plants.
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The data show that Tyr-37 is important for resveratrol IAPP interactions because mutation of this residue reduces the effect of the compound on IAPP amyloid formation.
The RB-mediated repression was dependent on E2F, because mutations of the E2F-binding sequences in the Cyclin A promoter abolished the RB-mediated transcription repression (not shown).
This enzyme is also suggested to be related to the antiviral immune response in humans, because mutations of the RNase H-encoding gene have been found in individuals affected by a human neurological disease, Aicardi-Goutières syndrome [ 20].
Thus it is unlikely that cluster N1a has a specific functional role in complex I. Instead, because mutations of the cluster ligands in both E. coli and Y. lipolytica preclude formation of a stable and functional enzyme, it is most likely that the correct incorporation of cluster N1a is required for enzyme assembly and stability.
The two branches are not redundant, because mutation of chico or the loss of its ability to activate either branch results in the same magnitude of lifespan extension.
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