6Q2M

Crystal structure of Photinus pyralis Luciferase Pps6 mutant in complex with DLSA

6Q2M の概要

• エントリーDOI: 10.2210/pdb6q2m/pdb
• 関連するPDBエントリー: 4G36
• 分子名称: Luciferin 4-monooxygenase, SULFATE ION, (2S,5S)-hexane-2,5-diol, ... (6 entities in total)
• 機能のキーワード: luciferase, adenylation domain, ligase, bioluminescence
• 由来する生物種: Photinus pyralis (Common eastern firefly)
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 188992.87

構造登録者

Patel, K.D.,Gulick, A.M. (登録日: 2019-08-08, 公開日: 2019-10-16, 最終更新日: 2023-10-11)

主引用文献

Branchini, B.R.,Fontaine, D.M.,Southworth, T.L.,Huta, B.P.,Racela, A.,Patel, K.D.,Gulick, A.M.
Mutagenesis and Structural Studies Reveal the Basis for the Activity and Stability Properties That Distinguish thePhotinusLuciferasesscintillansandpyralis.
Biochemistry, 58:4293-4303, 2019

PubMed Abstract: The dazzling yellow-green light emission of the common North American firefly and other bioluminescent organisms has provided a wide variety of prominent research applications like reporter gene assays and in vivo imaging methods. While the enzyme has been extensively studied, only recently has a second luciferase been cloned from the species . Even though the enzymes share very high sequence identity (89.8%), the color of the light they emit, their specific activity and their stability to heat, pH, and chemical denaturation are quite different with the luciferase being generally more resistant. Through the construction and evaluation of the properties of chimeric domain swapped, single point, and various combined variants, we have determined that only six amino acid changes are necessary to confer all of the properties of the enzyme to wild-type luciferase. Altered stability properties were attributed to four of the amino acid changes (T214N/S276T/H332N/E354N), and single mutations each predominantly changed emission color (Y255F) and specific activity (A222C). Results of a crystallographic study of the enzyme containing the six changes (Pps6) provide some insight into the structural basis for some of the documented property differences.

PubMed: 31560532
DOI: 10.1021/acs.biochem.9b00719

実験手法

X-RAY DIFFRACTION (2.75 Å)