PDBsum entry 4jef

Transferase

PDB id: 4jef

Contents

Protein chain

248 a.a.

Ligands

SO4 ×3

Waters ×24

PDB id:

4jef

Name:

Transferase

Title:

Crystal structure of human thymidylate synthase y202a in inactive conformation.

Structure:

Thymidylate synthase. Chain: a. Synonym: ts, tsase. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: tyms, ts, ok/sw-cl.29. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.31Å R-factor: 0.187 R-free: 0.222

Authors:

A.Tochowicz,R.M.Stroud,R.C.Wade

Key ref:

O.M.Salo-Ahen et al. (2015). Hotspots in an obligate homodimeric anticancer target. Structural and functional effects of interfacial mutations in human thymidylate synthase. J Med Chem, 58, 3572-3581. PubMed id: 25798950 DOI: 10.1021/acs.jmedchem.5b00137

Date:

26-Feb-13 Release date: 20-Aug-14

Protein chain

P04818  (TYSY_HUMAN) -  Thymidylate synthase from Homo sapiens

Seq: 313 a.a.

Struc: 248 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.1.1.45 - thymidylate synthase.
• Pathway: Folate Coenzymes
• Reaction: dUMP + (6R)-5,10-methylene-5,6,7,8-tetrahydrofolate = 7,8-dihydrofolate + dTMP

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

reference

DOI no: 10.1021/acs.jmedchem.5b00137

J Med Chem 58:3572-3581 (2015)

PubMed id: 25798950

Hotspots in an obligate homodimeric anticancer target. Structural and functional effects of interfacial mutations in human thymidylate synthase.

O.M.Salo-Ahen, A.Tochowicz, C.Pozzi, D.Cardinale, S.Ferrari, Y.Boum, S.Mangani, R.M.Stroud, P.Saxena, H.Myllykallio, M.P.Costi, G.Ponterini, R.C.Wade.

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.