2XZ7

CRYSTAL STRUCTURE OF THE PHOSPHOENOLPYRUVATE-BINDING DOMAIN OF ENZYME I IN COMPLEX WITH PHOSPHOENOLPYRUVATE FROM THE THERMOANAEROBACTER TENGCONGENSIS PEP-SUGAR PHOSPHOTRANSFERASE SYSTEM (PTS)

2XZ7 の概要

• エントリーDOI: 10.2210/pdb2xz7/pdb
• 関連するPDBエントリー: 2BG5 2XZ9
• 分子名称: PHOSPHOENOLPYRUVATE-PROTEIN KINASE (PTS SYSTEM EI COMPONENT IN BACTERIA), PHOSPHOENOLPYRUVATE, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: thermophilic, pep-utilising enzyme, transferase
• 由来する生物種: THERMOANAEROBACTER TENGCONGENSIS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36492.37

構造登録者

Waltersperger, S.M.,Oberholzer, A.E.,Schneider, P.,Baumann, U.,Erni, B. (登録日: 2010-11-23, 公開日: 2011-06-15, 最終更新日: 2023-12-20)

主引用文献

Navdaeva, V.,Zurbriggen, A.,Waltersperger, S.M.,Schneider, P.,Oberholzer, A.E.,Bahler, P.,Bachler, C.,Grieder, A.,Baumann, U.,Erni, B.
Phosphoenolpyruvate: Sugar Phosphotransferase System from the Hyperthermophilic Thermoanaerobacter Tengcongensis.
Biochemistry, 50:1184-, 2011

PubMed Abstract: Thermoanaerobacter tengcongensis is a thermophilic eubacterium that has a phosphoenolpyruvate (PEP) sugar phosphotransferase system (PTS) of 22 proteins. The general PTS proteins, enzyme I and HPr, and the transporters for N-acetylglucosamine (EIICB(GlcNAc)) and fructose (EIIBC(Fru)) have thermal unfolding transitions at ∼90 °C and a temperature optimum for in vitro sugar phosphotransferase activity of 65 °C. The phosphocysteine of a EIICB(GlcNAc) mutant is unusually stable at room temperature with a t(1/2) of 60 h. The PEP binding C-terminal domain of enzyme I (EIC) forms a metastable covalent adduct with PEP at 65 °C. Crystallization of this adduct afforded the 1.68 Å resolution structure of EIC with a molecule of pyruvate in the active site. We also report the 1.83 Å crystal structure of the EIC-PEP complex. The comparison of the two structures with the apo form and with full-length EI shows differences between the active site side chain conformations of the PEP and pyruvate states but not between the pyruvate and apo states. In the presence of PEP, Arg465 forms a salt bridge with the phosphate moiety while Glu504 forms salt bridges with Arg186 and Arg195 of the N-terminal domain of enzyme I (EIN), which stabilizes a conformation appropriate for the in-line transfer of the phosphoryl moiety from PEP to His191. After transfer, Arg465 swings 4.8 Å away to form an alternative salt bridge with the carboxylate of Glu504. Glu504 loses the grip of Arg186 and Arg195, and the EIN domain can swing away to hand on the phosphoryl group to the phosphoryl carrier protein HPr.

PubMed: 21250658
DOI: 10.1021/BI101721F

実験手法

X-RAY DIFFRACTION (1.83 Å)