1VZA

THYMIDYLATE SYNTHASE E60D MUTANT BINARY COMPLEX WITH 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (DUMP)

1VZA の概要

• エントリーDOI: 10.2210/pdb1vza/pdb
• 分子名称: THYMIDYLATE SYNTHASE, 2'-DEOXYURIDINE 5'-MONOPHOSPHATE (3 entities in total)
• 機能のキーワード: methyltransferase, nucleotide synthase
• 由来する生物種: Lactobacillus casei
• 細胞内の位置: Cytoplasm: P00469
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36880.60

構造登録者

Birdsall, D.L.,Finer-Moore, J.,Stroud, R.M. (登録日: 1996-09-18, 公開日: 1997-03-12, 最終更新日: 2024-02-14)

主引用文献

Birdsall, D.L.,Huang, W.,Santi, D.V.,Stroud, R.M.,Finer-Moore, J.
The separate effects of E60Q in Lactobacillus casei thymidylate synthase delineate between mechanisms for formation of intermediates in catalysis.
Protein Eng., 11:171-183, 1998

PubMed Abstract: X-Ray crystal structures of Lactobacillus casei thymidylate synthase (TS) mutant complexes of E60D with dUMP, and E60Q with dUMP or FdUMP, as well as ternary complexes with folate analog inhibitor CB3717, are described. The structures we report address the decrease in rate of formation of ternary complexes in the E60 mutants. Structures of ternary complexes of L.casei TS mimic ligand-bound TS just prior to covalent bond formation between ligands and protein. Ternary complex structures of L.casei TS E60Q show the ligands are not optimally aligned for making the necessary covalent bonds. Since CB3717 is an analog of the open, activated form of the cofactor, these structures suggest that the slow rate of ternary complex formation in E60 mutants is at least partly the result of impaired alignment of ligands in the active site after binding and activation of the cofactor. Binary complexes of TS E60Q and TS E60D with substrate (dUMP) show no change in dUMP position or occupancy. These results are consistent with the fact that Kd(dUMP) and Km(dUMP) are almost the same, and the rates of folate-independent debromination of 5-bromo-dUMP are even higher than for wild type TS.

PubMed: 9613841
DOI: 10.1093/protein/11.3.171

実験手法

X-RAY DIFFRACTION (2.5 Å)