4KPW
Crystal structure of His-tagged human thymidylate synthase R175A mutant
Summary for 4KPW
| Entry DOI | 10.2210/pdb4kpw/pdb |
| Related | 3N5E 3N5G |
| Descriptor | Thymidylate synthase, SULFATE ION, 1,2-ETHANEDIOL, ... (4 entities in total) |
| Functional Keywords | interface hot spot, transferase, methyltransferase, nucleotide biosynthesis |
| Biological source | Homo sapiens (human) |
| Cellular location | Nucleus : P04818 |
| Total number of polymer chains | 1 |
| Total formula weight | 37831.17 |
| Authors | Pozzi, C.,Mangani, S. (deposition date: 2013-05-14, release date: 2014-05-14, Last modification date: 2023-09-20) |
| Primary citation | Salo-Ahen, O.M.,Tochowicz, A.,Pozzi, C.,Cardinale, D.,Ferrari, S.,Boum, Y.,Mangani, S.,Stroud, R.M.,Saxena, P.,Myllykallio, H.,Costi, M.P.,Ponterini, G.,Wade, R.C. Hotspots in an obligate homodimeric anticancer target. Structural and functional effects of interfacial mutations in human thymidylate synthase. J.Med.Chem., 58:3572-3581, 2015 Cited by PubMed Abstract: Human thymidylate synthase (hTS), a target for antiproliferative drugs, is an obligate homodimer. Single-point mutations to alanine at the monomer-monomer interface may enable the identification of specific residues that delineate sites for drugs aimed at perturbing the protein-protein interactions critical for activity. We computationally identified putative hotspot residues at the interface and designed mutants to perturb the intersubunit interaction. Dimer dissociation constants measured by a FRET-based assay range from 60 nM for wild-type hTS up to about 1 mM for single-point mutants and agree with computational predictions of the effects of these mutations. Mutations that are remote from the active site retain full or partial activity, although the substrate KM values were generally higher and the dimer was less stable. The lower dimer stability of the mutants can facilitate access to the dimer interface by small molecules and thereby aid the design of inhibitors that bind at the dimer interface. PubMed: 25798950DOI: 10.1021/acs.jmedchem.5b00137 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.03 Å) |
Structure validation
Download full validation report
