1E2P

Thymidine kinase, DHBT

1E2P の概要

• エントリーDOI: 10.2210/pdb1e2p/pdb
• 関連するPDBエントリー: 1E2H 1E2I 1E2J 1E2K 1E2L 1E2M 1E2N 1KI2 1KI4 1KI5 1KI6 1KI7 1KI8 1KIM 1VTK 2VTK 3VTK
• 分子名称: THYMIDINE KINASE, SULFATE ION, 6-[3-HYDROXY-2-(HYDROXYMETHYL)PROPYL]-5-METHYL-2,4(1H,3H)-PYRIMIDINEDIONE, ... (4 entities in total)
• 機能のキーワード: transferase, kinase, dna synthesis, atp-binding
• 由来する生物種: HERPES SIMPLEX VIRUS TYPE 1
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 72178.73

構造登録者

Schulz, G.E.,Kessler, U. (登録日: 2000-05-23, 公開日: 2001-04-26, 最終更新日: 2023-12-06)

主引用文献

Wurth, C.,Kessler, U.,Vogt, J.,Schulz, G.E.,Folkers, G.,Scapozza, L.
The Effect of Substrate Binding on the Conformation and Structural Stability of Herpes Simplex Virus Type 1 Thymidine Kinase
Protein Sci., 10:63-, 2001

PubMed Abstract: The structure of Herpes simplex virus type 1 thymidine kinase (TK(HSV1)) is known at high resolution in complex with a series of ligands and exhibits important structural similarities to the nucleoside monophosphate (NMP) kinase family, which are known to show large conformational changes upon binding of substrates. The effect of substrate binding on the conformation and structural stability of TK(HSV1), measured by thermal denaturation experiments, far-UV circular dichroism (CD) and fluorescence is described, and the results indicate that the conformation of the ligand-free TK(HSV1) is less ordered and less stable compared to the ligated enzyme. Furthermore, two crystal structures of TK(HSV1) in complex with two new ligands, HPT and HMTT, refined to 2.2 A are presented. Although TK(HSV1):HPT does not exhibit any significant deviations from the model of TK(HSV1):dT, the TK(HSV1):HMTT complex displays a unique conformationally altered active site resulting in a lowered thermal stability of this complex. Moreover, we show that binding affinity and binding mode of the ligand correlate with thermal stability of the complex. We use this correlation to propose a method to estimate binding constants for new TK(HSV1)substrates using thermal denaturation measurements monitored by CD spectroscopy. The kinetic and structural results of both test substrates HPT and HMTT show that the CD thermal denaturation system is very sensitive to conformational changes caused by unusual binding of a substrate analog.

PubMed: 11266595
DOI: 10.1110/PS.27401

実験手法

X-RAY DIFFRACTION (2.5 Å)