PDBsum entry 5os2

Transferase

PDB id: 5os2

Contents

Protein chain

261 a.a.

Ligands

ADP

A7K

Metals

_MG ×2

Waters ×123

PDB id:

5os2

Name:

Transferase

Title:

Crystal structure of aurora-a kinase in complex with an allosterically binding fragment

Structure:

Aurora kinase a. Chain: a. Synonym: aurora 2,aurora/ipl1-related kinase 1,hark1,breast tumor- amplified kinase,serine/threonine-protein kinase 15,serine/threonine- protein kinase 6,serine/threonine-protein kinase aurora-a. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: aurka, aik, airk1, ark1, aura, ayk1, btak, iak1, stk15, stk6. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.92Å R-factor: 0.213 R-free: 0.251

Authors:

P.J.Mcintyre,P.M.Collins,F.Von Delft,R.Bayliss

Key ref:

P.J.McIntyre et al. (2017). Characterization of Three Druggable Hot-Spots in the Aurora-A/TPX2 Interaction Using Biochemical, Biophysical, and Fragment-Based Approaches. ACS Chem Biol, 12, 2906-2914. PubMed id: 29045126 DOI: 10.1021/acschembio.7b00537

Date:

16-Aug-17 Release date: 01-Nov-17

Protein chain

O14965  (AURKA_HUMAN) -  Aurora kinase A from Homo sapiens

Seq: 403 a.a.

Struc: 261 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.7.11.1 - non-specific serine/threonine protein kinase.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] (Bound ligand (Het Group name = ADP) corresponds exactly) + ADP + H(+)

L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] (Bound ligand (Het Group name = ADP) corresponds exactly) + ADP + H(+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1021/acschembio.7b00537

ACS Chem Biol 12:2906-2914 (2017)

PubMed id: 29045126

Characterization of Three Druggable Hot-Spots in the Aurora-A/TPX2 Interaction Using Biochemical, Biophysical, and Fragment-Based Approaches.

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.

ABSTRACT

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.