Počet záznamů: 1
Characterization of Three Druggable Hot-Spots in the Aurora-A/TPX2 Interaction Using Biochemical, Biophysical, and Fragment-Based Approaches
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SYSNO ASEP 0485445 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Characterization of Three Druggable Hot-Spots in the Aurora-A/TPX2 Interaction Using Biochemical, Biophysical, and Fragment-Based Approaches Tvůrce(i) McIntyre, P. J. (GB)
Collins, P. M. (GB)
Vrzal, Lukáš (UOCHB-X)
Birchall, K. (GB)
Arnold, L. H. (GB)
Mpamhanga, C. (GB)
Coombs, P. J. (GB)
Burgess, S. G. (GB)
Richards, M. W. (GB)
Winter, A. (GB)
Veverka, Václav (UOCHB-X) RID, ORCID
von Delft, F. (GB)
Merritt, A. (GB)
Bayliss, R. (GB)Zdroj.dok. ACS Chemical Biology. - : American Chemical Society - ISSN 1554-8929
Roč. 12, č. 11 (2017), s. 2906-2914Poč.str. 9 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova fragment-based drug discovery ; structural biology ; inhibition Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP LK11205 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy LO1304 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora UOCHB-X - RVO:61388963 UT WOS 000416204500026 EID SCOPUS 85034666028 DOI https://doi.org/10.1021/acschembio.7b00537 Anotace 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. Pracoviště Ústav organické chemie a biochemie Kontakt asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Rok sběru 2018
Počet záznamů: 1