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Figure 2.
(a) NMR chemical shift perturbations induced by binding of
ClipZn12, mapped onto the surface of ClipCG1 (left; PDB 2CP7)
and ClipCG2 (right; PDB 2CP6). The two panels show equivalent
views. Purple residues are missing from the HSQC spectra owing
to chemical exchanges. See Supplementary Figure 2. (b) Sequence
alignments of ClipCG domains. ClipCG1 and ClipCG2 of human,
Xenopus laevis and zebrafish CLIP170, human CLIP115 and
Drosophila CLIP190 are shown. The single ClipCG domains of
p150n, fission yeast Tip1p and budding yeast Bik1 are also
shown. Green highlight, GKNDG motif; blue, its invariant lysine
residue; red, arginine residues conserved in ClipCG1 (and
p150n); orange, lysine and histidine conserved in ClipCG2. (c,d)
In vitro assays of wild-type or mutant ClipCG1 and ClipCG2
binding to ClipZn12 (c; see Supplementary Table 2) or MTs (d).
ClipCG1, ClipCG2 and mutants were detected by Coomassie staining
after SDS-PAGE. Input lanes show wild-type ClipCG domains, for
reference. All the mutations abrogate binding to ClipZn12 or MTs.
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