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PDBsum entry 2hqh
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Structural protein, protein binding
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PDB id
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2hqh
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References listed in PDB file
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Key reference
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Title
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Clip170 autoinhibition mimics intermolecular interactions with p150glued or eb1.
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Authors
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I.Hayashi,
M.J.Plevin,
M.Ikura.
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Ref.
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Nat Struct Biol, 2007,
14,
980-981.
[DOI no: ]
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PubMed id
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Abstract
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CLIP170 and p150(Glued) localize to the plus ends of growing microtubules. Using
crystallography and NMR, we show that autoinhibitory interactions within CLIP170
use the same binding determinants as CLIP170's intermolecular interactions with
p150(Glued). These interactions have both similar and distinct features when
compared with the p150(Glued)-EB1 complex. Our data thus demonstrate that
regulation of microtubule dynamics by plus end-tracking proteins (+TIPs) occurs
through direct competition between homologous binding interfaces.
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Figure 1.
(a) Surface representation of p150n with C  traces
of ClipZn2 (pink) and p150n (green). Key interacting side chains
are shown as sticks (green atoms are sulfur). Blue, GKNDG motif;
orange, conserved hydrophobic residues (Phe52, Trp57 and Phe88;
Supplementary Fig. 3 online). Red circled region is expanded in
b. (b) Close-up views of interface between ClipZn2 and p150n.
Middle panel has same orientation as in a. Residues discussed in
the text are labeled. Orange sphere, water molecule mediating
the Arg90-Asn1422 interaction. (c) Pull-down assays of wild-type
and mutant ClipZn12 and p150n. p150n proteins were incubated
with glutathione S-transferase (GST)-fused ClipZn12 proteins.
Left lane contains wild-type p150n, for reference. Mock
pull-down is shown as a control (see Supplementary Table 2).
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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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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nat Struct Biol
(2007,
14,
980-981)
copyright 2007.
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