2ZO6

Crystal Structure of Kusabira-Cyan (KCY), a Cyan-Emitting GFP-Like Protein

「2EJH」から置き換えられました

2ZO6 の概要

• エントリーDOI: 10.2210/pdb2zo6/pdb
• 関連するPDBエントリー: 2ZO7
• 分子名称: CYAN-EMITTING GFP-LIKE PROTEIN, KUSABIRA-CYAN (KCY) (2 entities in total)
• 機能のキーワード: gfp-like protein, luminescent protein, structural genomics, riken structural genomics/proteomics initiative, rsgi, nppsfa, national project on protein structural and functional analyses
• 由来する生物種: Fungia concinna (Mushroom coral)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28224.95

構造登録者

Kikuchi, A.,Fukumura, E.,Karasawa, S.,Miyawaki, A.,Shiro, Y.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (登録日: 2008-05-06, 公開日: 2009-05-12, 最終更新日: 2024-10-16)

主引用文献

Kikuchi, A.,Fukumura, E.,Karasawa, S.,Shiro, Y.,Miyawaki, A.
Crystal structure of a new cyan fluorescent protein and its hue-shifted variants
Biochemistry, 48:5276-5283, 2009

PubMed Abstract: Green fluorescent protein (GFP) based techniques are well established in molecular biology; however, the detailed mechanism for the fine-tuning of fluorescent colors remains unclear. Here, we report the cloning and crystal structure of a new cyan-emitting GFP-like protein, KCy. We also developed a mutant protein with a high folding efficiency (KCy-G4219: lambda(abs) = 453 nm; lambda(em) = 486 nm). X-ray diffraction analysis revealed that the KCy chromophore is formed from an internal Ser62-Tyr63-Gly64 tripeptide. The serine residue at the first position of the chromophore-forming tripeptide has a short polar chain (-OH) that forms a noncovalent interaction with the His38 imidazole at a distance of 2.96 A. Substitution of His38 in KCy-G4219 with Gln (KCy-R1) or Leu residues resulted in a slight but significant red shift of the emission peak maximum from 486 to 492 or 496 nm, respectively. The crystal structure of KCy-R1 determined at a resolution of 1.58 A showed that the noncovalent interaction between Ser62-OH and the substituted Gln38 occurred over a longer distance (3.07 A) than that observed in the wild-type KCy. Such an interaction is absent in the Leu mutant, suggesting that this interaction is one of the key factors responsible for fine-tuning the emission peak maxima, which are affected by chromophore polarization. Moreover, the structural comparison suggests that an additional water molecule buried in the space between the Ala158 residue and the chromophore phenolate is also responsible for the chromophore polarization.

PubMed: 19402703
DOI: 10.1021/bi801658p

実験手法

X-RAY DIFFRACTION (1.4 Å)