title: MTH1 Substrate Recognition--An Example of Specific Promiscuity.
abstract: MTH1 (NUDT1) is an oncologic target involved in the prevention of DNA damage. We investigate the way MTH1 recognises its substrates and present substrate-bound structures of MTH1 for 8-oxo-dGTP and 8-oxo-rATP as examples of novel strong and weak binding substrate motifs. Investigation of a small set of purine-like fragments using 2D NMR resulted in identification of a fragment with weak potency. The protein-ligand X-Ray structure of this fragment provides insight into the role of water molecules in substrate selectivity. Wider fragment screening by NMR resulted in three new protein structures exhibiting alternative binding configurations to the key Asp-Asp recognition element of the protein. These inhibitor binding modes demonstrate that MTH1 employs an intricate yet promiscuous mechanism of substrate anchoring through its Asp-Asp pharmacophore. The structures suggest that water-mediated interactions convey selectivity towards oxidized substrates over their non-oxidised counterparts, in particular by stabilization of a water molecule in a hydrophobic environment through hydrogen bonding. These findings may be useful in the design of inhibitors of MTH1.
authors.affiliation: Chemistry, Oncology, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom, and Alderley Park, Cheshire, SK10 4TG, United Kingdom.
authors.forename: J Willem M
authors.initials: JW
authors.lastname: Nissink
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom.
authors.forename: Michal
authors.initials: M
authors.lastname: Bista
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom.
authors.forename: Jason
authors.initials: J
authors.lastname: Breed
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom.
authors.forename: Nikki
authors.initials: N
authors.lastname: Carter
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, United Kingdom.
authors.forename: Kevin
authors.initials: K
authors.lastname: Embrey
authors.affiliation: Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Read
authors.affiliation: Chemistry, Oncology, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Unit 310 (Darwin Building), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom, and Alderley Park, Cheshire, SK10 4TG, United Kingdom.
authors.forename: Jon J
authors.initials: JJ
authors.lastname: Winter-Holt
mesh_terms: D020816:Amino Acid Motifs; D001224:Aspartic Acid / Q000378:metabolism; D001665:Binding Sites; D018360:Crystallography, X-Ray; D003848:Deoxyguanine Nucleotides / Q000737:chemistry / Q000378:metabolism; D006860:Hydrogen Bonding; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D017433:Protein Structure, Secondary; D013379:Substrate Specificity; D014867:Water
chemical_list: D003848:Deoxyguanine Nucleotides; D014867:Water; C078206:8-oxodeoxyguanosine triphosphate; D001224:Aspartic Acid; D010744:Phosphoric Monoester Hydrolases