Exact(12)
The STDD mutant did not obviously affect their interactions (Fig. 2D), but T206D and Y217D could abrogate the interaction of EB1 with CLIP170 and p150Glued (Fig. 2D).
Considering that phosphorylation near the SxIP motif can abrogate the interaction between EB1 and +TIPs (Honnappa et al., 2009), we speculated that amino acids with negative charge (D or E) near the SxIP motif might be responsible for the different abilities of EB1 to interact with different +TIPs (Fig. 2C).
In line with this interpretation, the P4270L mutation is able to completely abrogate the interaction with NPHP1-SH3 both in GST pull-down assays and in the co-immunoprecipitation studies.
Accordingly, the Δ602 630 mutation should discontinue the substrate-binding cleft and abrogate the interaction with myosin.
In contrast, K45A and R47A did not abrogate the interaction, and the F51A mutation disrupts the structure of CYP33 RRM (data not shown).
However, in this case, GW2974 reduced, but did not completely abrogate the interaction (compare lanes 7 8 with lanes 3 4).
Similar(48)
We then demonstrate that the Dnmt1 phosphorylations by Akt and/or PKC abrogate the interactions of Dnmt1 with PCNA and UHRF1 in cellular and acelluar studies including mass spectrometric analyses and the use of primary cultured patient-derived glioma.
Indeed, a single nucleotide mutation in Unc93b1 gene (Unc93b1 3d/3d) abrogates the interaction with TLR3, 7, and 9, but also cross-presentation of antigens by MHCI molecules, even in the absence of TLR help19.
Additionally, we showed that a mutation in the Ca2+ EF-hand motif (STIM1-D76A), that renders STIM1 constitutively active, abrogates the interaction with UNC93B1, suggesting that the association between the two proteins requires STIM1 Ca2+-binding domain.
Unexpectedly, Y217F also abrogated the interaction of EB1 with MCAK and APC (Fig. 2A).
Y217D, but not T206D or STDD, abrogated the interaction of EB1 with MCAK and APC (Fig. 2A).
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