1MYW

CRYSTAL STRUCTURE OF A YELLOW FLUORESCENT PROTEIN WITH IMPROVED MATURATION AND REDUCED ENVIRONMENTAL SENSITIVITY

1MYW の概要

• エントリーDOI: 10.2210/pdb1myw/pdb
• 分子名称: Green fluorescent protein (2 entities in total)
• 機能のキーワード: bioluminescence, photoactive protein, green fluorescent protein, yellow-emission variant, improved maturation, beta-barrel, chromophore, luminescent protein
• 由来する生物種: Aequorea victoria
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 26875.29

構造登録者

Rekas, A.,Alattia, J.R.,Nagai, T.,Miyawaki, A.,Ikura, M. (登録日: 2002-10-04, 公開日: 2003-01-14, 最終更新日: 2026-03-18)

主引用文献

Rekas, A.,Alattia, J.R.,Nagai, T.,Miyawaki, A.,Ikura, M.
Crystal Structure of Venus, a Yellow Fluorescent Protein with Improved Maturation and Reduced Environmental Sensitivity
J.Biol.Chem., 277:50573-50578, 2002

PubMed Abstract: Yellow emission variants of green fluorescent protein (GFP) have been found useful in a variety of applications in biological systems due to their red-shifted emission spectrum and sensitivity to environmental parameters, such as pH and ionic strength. However, slow maturation properties and new requirements for more intense fluorescence necessitated further mutagenesis studies of these proteins. Venus, a new variant with improved maturation and brightness, as well as reduced environmental dependence, was recently developed by introducing five mutations into the well characterized variant, enhanced yellow fluorescent protein (EYFP). In this paper, we present the crystal structure of Venus at 2.2 A resolution, which enabled us to correlate its novel features with these mutation points. The rearrangement of several side chains near the chromophore, initiated by the F46L mutation, was found to improve maturation at 37 degrees C by removing steric and energetic constraints, which may hinder folding of the polypeptide chain, and by accelerating the oxidation of the Calpha-Cbeta bond of Tyr(66) during chromophore formation. M153T, V163A, and S175G were also found to improve the rate of maturation by creating regions of greater flexibility. F64L induced large conformational changes in the molecule, leading to the removal of halide sensitivity by preventing ion access to the binding site.

PubMed: 12370172
DOI: 10.1074/jbc.M209524200

実験手法

X-RAY DIFFRACTION (2.2 Å)