4J2P

Crystal structure of LuxF from Photobacterium leiognathi

4J2P の概要

• エントリーDOI: 10.2210/pdb4j2p/pdb
• 関連するPDBエントリー: 1NFP
• 分子名称: Non-fluorescent flavoprotein (2 entities in total)
• 機能のキーワード: incomplete beta-barrel, modified 7-stranded barrel, myr-fmn binding, myr-fmn, luminescent protein
• 由来する生物種: Photobacterium leiognathi
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27340.82

構造登録者

Winkler, A.,Macheroux, P.,Gruber, K. (登録日: 2013-02-05, 公開日: 2014-02-05, 最終更新日: 2023-09-20)

主引用文献

Bergner, T.,Tabib, C.R.,Winkler, A.,Stipsits, S.,Kayer, H.,Lee, J.,Malthouse, J.P.,Mayhew, S.,Muller, F.,Gruber, K.,Macheroux, P.
Structural and biochemical properties of LuxF from Photobacterium leiognathi.
Biochim. Biophys. Acta, 1854:1466-1475, 2015

PubMed Abstract: The lux-operon of bioluminescent bacteria contains the genes coding for the enzymes required for light emission. Some species of Photobacteria feature an additional gene, luxF, which shows similarity to luxA and luxB, the genes encoding the heterodimeric luciferase. Isolated dimeric LuxF binds four molecules of an unusually derivatized flavin, i.e., 6-(3'-(R)-myristyl)-FMN (myrFMN). In the present study we have heterologously expressed LuxF in Escherichia coli BL21 in order to advance our understanding of the protein's binding properties and its role in photobacterial bioluminescence. Structure determination by X-ray crystallography confirmed that apo-LuxF possesses four preorganized binding sites, which are further optimized by adjusting the orientation of amino acid side chains. To investigate the binding properties of recombinant LuxF we have isolated myrFMN from Photobacterium leiognathi S1. We found that LuxF binds myrFMN tightly with a dissociation constant of 80±20 nM demonstrating that the purified apo-form of LuxF is fully competent in myrFMN binding. In contrast to LuxF, binding of myrFMN to luciferase is much weaker (Kd=4.0±0.4 μM) enabling LuxF to prevent inhibition of the enzyme by scavenging myrFMN. Moreover, we have used apo-LuxF to demonstrate that myrFMN occurs in all Photobacteria tested, irrespective of the presence of luxF indicating that LuxF is not required for myrFMN biosynthesis.

PubMed: 26209460
DOI: 10.1016/j.bbapap.2015.07.008

実験手法

X-RAY DIFFRACTION (1.85 Å)