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PDBsum entry 5os2
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References listed in PDB file
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Key reference
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Title
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Characterization of three druggable hot-Spots in the aurora-A/tpx2 interaction using biochemical, Biophysical, And fragment-Based approaches.
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Authors
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P.J.Mcintyre,
P.M.Collins,
L.Vrzal,
K.Birchall,
L.H.Arnold,
C.Mpamhanga,
P.J.Coombs,
S.G.Burgess,
M.W.Richards,
A.Winter,
V.Veverka,
F.V.Delft,
A.Merritt,
R.Bayliss.
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Ref.
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ACS Chem Biol, 2017,
12,
2906-2914.
[DOI no: ]
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PubMed id
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Abstract
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The mitotic kinase Aurora-A and its partner protein TPX2 (Targeting Protein for
Xenopus kinesin-like protein 2) are overexpressed in cancers, and it has been
proposed that they work together as an oncogenic holoenzyme. TPX2 is responsible
for activating Aurora-A during mitosis, ensuring proper cell division.
Disruption of the interface with TPX2 is therefore a potential target for novel
anticancer drugs that exploit the increased sensitivity of cancer cells to
mitotic stress. Here, we investigate the interface using coprecipitation assays
and isothermal titration calorimetry to quantify the energetic contribution of
individual residues of TPX2. Residues Tyr8, Tyr10, Phe16, and Trp34 of TPX2 are
shown to be crucial for robust complex formation, suggesting that the
interaction could be abrogated through blocking any of the three pockets on
Aurora-A that complement these residues. Phosphorylation of Aurora-A on Thr288
is also necessary for high-affinity binding, and here we identify arginine
residues that communicate the phosphorylation of Thr288 to the TPX2 binding
site. With these findings in mind, we conducted a high-throughput X-ray
crystallography-based screen of 1255 fragments against Aurora-A and identified
59 hits. Over three-quarters of these hits bound to the pockets described above,
both validating our identification of hotspots and demonstrating the
druggability of this protein-protein interaction. Our study exemplifies the
potential of high-throughput crystallography facilities such as XChem to aid
drug discovery. These results will accelerate the development of chemical
inhibitors of the Aurora-A/TPX2 interaction.
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