6R2E

Crystal structure of the human thymidylate synthase (hTS) interface variant Q62R

6R2E の概要

• エントリーDOI: 10.2210/pdb6r2e/pdb
• 分子名称: Thymidylate synthase, SULFATE ION, N-[4-({[(6S)-2-amino-5-formyl-4-oxo-3,4,5,6,7,8-hexahydropteridin-6-yl]methyl}amino)benzoyl]-L-glutamic acid, ... (7 entities in total)
• 機能のキーワード: human thymidylate synthase, folate pathway, q62r, interface variant, transferase
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 304626.38

構造登録者

Pozzi, C.,Mangani, M. (登録日: 2019-03-16, 公開日: 2019-04-10, 最終更新日: 2024-01-24)

主引用文献

Pozzi, C.,Lopresti, L.,Santucci, M.,Costi, M.P.,Mangani, S.
Evidence of Destabilization of the Human Thymidylate Synthase (hTS) Dimeric Structure Induced by the Interface Mutation Q62R.
Biomolecules, 9:-, 2019

PubMed Abstract: In human cells, thymidylate synthase (TS) provides the only source of 2'-deoxythymidyne-5'-monophosphate (dTMP), which is required for DNA biosynthesis. Because of its pivotal role, human TS (hTS) represents a validated target for anticancer chemotherapy. Nonetheless, the efficacy of drugs blocking the hTS active site has limitations due to the onset of resistance in cancer cells, requiring the identification of new strategies to effectively inhibit this enzyme. Human TS works as an obligate homodimer, making the inter-subunit interface an attractive targetable area. Here, we report the design and investigation of a new hTS variant, in which Gln62, located at the dimer interface, has been replaced by arginine in order to destabilize the enzyme quaternary assembly. The hTS Q62R variant has been characterized though kinetic assay, thermal denaturation analysis and X-ray crystallography. Our results provide evidence that hTS Q62R has a reduced melting temperature. The effective destabilization of the TS quaternary structure is also confirmed by structural analysis, showing that the introduced mutation induces a slight aperture of the hTS dimer. The generation of hTS variants having a more accessible interface area can facilitate the screening of interface-targeting molecules, providing key information for the rational design of innovative hTS interface inhibitors.

PubMed: 30987202
DOI: 10.3390/biom9040134

実験手法

X-RAY DIFFRACTION (2.55 Å)