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As shown in Fig. 5A, the K189A rNSs mutant was completely inactive whereas the D159A mutant was partially active.
As in the cAMP assay, the MT1-I49N mutant was completely inactive in the ERK1/2 assay (Figure 3C).
Indeed, the GTPase activity of N169Q mutant was completely abolished and could not be measured (Table 2).
Furthermore, in Anabaena, a patB deletion mutant was completely defective for diazotrophic growth, but in the wildtype, its expression was restricted to heterocysts [41].
At both temperatures, the prkaca-F327A mutant was completely non-viable and had very low steady-state protein expression (Figure 3A).
While W542L mutant was completely devoid of enzyme activity, modifying the W542 residue with other amino acid residues partially recovered the TPPI activity.
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Interestingly, when the two mutations K98A and R185A are combined, the resulting APE1 double mutant is completely devoid of its DHU-, αdA-NIR and 3'→5' exonuclease activities, indicating that these two residues are indispensable for the NIR and exonuclease functions.
This correlates well with the fact that the dually phosphorylated PPPSPXS motif upon transferred to a heterologous receptor is sufficient to activate β-catenin signaling in vivo, whereas the PPPAPXA mutant is completely inactive [34], [35], [37].
Therefore, the mutant is completely sterile.
In vitro, we have shown that indeed the P56S MSP mutant is completely insoluble in buffers.
The Δ21 24 deletion mutant is completely unable to bind to pRb and does not induce tetrasomy.
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