4KH6

Toxoplasma gondii NTPDase1 C258S/C268S E493G crystallized with Mg and AMPNP

4KH6 の概要

• エントリーDOI: 10.2210/pdb4kh6/pdb
• 関連するPDBエントリー: 4A57 4A59 4A5A 4A5B 4JEP 4KH4 4KH5
• 分子名称: Nucleoside-triphosphatase 2, 5'-O-[(R)-hydroxy(phosphonoamino)phosphoryl]adenosine, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: hydrolase, actin-like fold, ntpdase
• 由来する生物種: Toxoplasma gondii
• 細胞内の位置: Secreted: Q27895
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 136845.07

構造登録者

Krug, U.,Totzauer, R.,Strater, N. (登録日: 2013-04-30, 公開日: 2013-11-06, 最終更新日: 2024-11-06)

主引用文献

Krug, U.,Totzauer, R.,Zebisch, M.,Strater, N.
The ATP/ADP substrate specificity switch between Toxoplasma gondii NTPDase1 and NTPDase3 is caused by an altered mode of binding of the substrate base.
Chembiochem, 14:2292-2300, 2013

PubMed Abstract: Two nucleoside triphosphate diphosphohydrolase isoforms (NTPDase1 and NTPDase3) are responsible for the hydrolysis of nucleotides by the intracellular protozoan Toxoplasma gondii. They constitute about 3 % of the total parasite protein. Despite sharing 97 % sequence identity they exhibit opposite ATP versus ADP substrate discrimination ratios. Here we show by mutagenesis that the residues G492/G493 in NTPDase3 and R492/E493 in NTPDase1 are predominantly responsible for the differences in substrate specificity. Crystal structures of NTPDase1 in complexation with analogues of ATP and ADP reveal that the inverted substrate specificity of NTPDase1 relative to NTPDase3 is achieved by switching from the canonical substrate binding mode to a very different alternative one. Instead of being stacked on top of a helix of the C-terminal domain the nucleotide base is positioned in the interdomain space between the side chains of R108 and R492, recruited from both domains. Furthermore, we show that the NTPDase1 substrate specificity is mainly dependent on the presence of the side chain of E493, which causes repositioning of the ribose component of the nucleotide. All in all, binding by the flexible side chains in the alternative binding mode in NTPDase1 allows for equally good positioning of ATP and ADP with increased activity toward ADP relative to what is seen in the case of NTPDase3.

PubMed: 24115522
DOI: 10.1002/cbic.201300441

実験手法

X-RAY DIFFRACTION (2.4 Å)