5YGG

Crystal structure of Fructokinase Double-Mutant (T261C-H108C) from Vibrio cholerae O395 in fructose, ADP and potassium ion bound form

5YGG の概要

• エントリーDOI: 10.2210/pdb5ygg/pdb
• 関連するPDBエントリー: 5EY7 5EYN 5F0Z 5F11
• 分子名称: Fructokinase, ADENOSINE-5'-DIPHOSPHATE, beta-D-fructofuranose, ... (6 entities in total)
• 機能のキーワード: kinase, sugar binding, double mutant, transferase
• 由来する生物種: Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 35649.09

構造登録者

Chatterjee, S.,Paul, R.,Nath, S.,Sen, U. (登録日: 2017-09-22, 公開日: 2018-09-26, 最終更新日: 2024-10-23)

主引用文献

Paul, R.,Chatterjee, S.,Nath, S.,Sen, U.
Large-scale conformational changes and redistribution of surface negative charge upon sugar binding dictate the fidelity of phosphorylation in Vibrio cholerae fructokinase.
Sci Rep, 8:16925-16925, 2018

PubMed Abstract: Fructokinase (FRK) catalyzes the first step of fructose metabolism i.e., D-fructose to D-fructose-6-phosphate (F6P), however, the mechanistic insights of this reaction are elusive yet. Here we demonstrate that the putative Vibrio cholerae fructokinase (VcFRK) exhibit strong fructose-6-kinase activity allosterically modulated by K/Cs. We have determined the crystal structures of apo-VcFRK and its complex with fructose, fructose-ADP-Ca, fructose-ADP-Ca-BeF. Collectively, we propose the catalytic mechanism and allosteric activation of VcFRK in atomistic details explaining why K/Cs are better activator than Na. Structural results suggest that apo VcFRK allows entry of fructose in the active site, sequester it through several conserved H-bonds and attains a closed form through large scale conformational changes. A double mutant (H108C/T261C-VcFRK), that arrests the closed form but unable to reopen for F6P release, is catalytically impotent highlighting the essentiality of this conformational change. Negative charge accumulation around ATP upon fructose binding, is presumed to redirect the γ-phosphate towards fructose for efficient phosphotransfer. Reduced phosphotransfer rate of the mutants E205Q and E110Q supports this view. Atomic resolution structure of VcFRK-fructose-ADP-Ca-BeF, reported first time for any sugar kinase, suggests that BeF moiety alongwith R176, Ca and 'anion hole' limit the conformational space for γ-phosphate favoring in-line phospho-transfer.

PubMed: 30446722
DOI: 10.1038/s41598-018-35236-3

実験手法

X-RAY DIFFRACTION (1.671 Å)