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2HGD

Structure of S65A Y66F GFP variant with an oxidized chromophore

2HGD の概要
エントリーDOI10.2210/pdb2hgd/pdb
関連するPDBエントリー2HGY 2hcg
分子名称Green fluorescent protein (2 entities in total)
機能のキーワードpost-translational modification, cyclization, oxidation, fluorophore, gfp, 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26825.14
構造登録者
Barondeau, D.P.,Kassmann, C.J.,Tainer, J.A.,Getzoff, E.D. (登録日: 2006-06-26, 公開日: 2007-03-27, 最終更新日: 2023-11-15)
主引用文献Barondeau, D.P.,Kassmann, C.J.,Tainer, J.A.,Getzoff, E.D.
The Case of the Missing Ring: Radical Cleavage of a Carbon-Carbon Bond and Implications for GFP Chromophore Biosynthesis
J.Am.Chem.Soc., 129:3118-3126, 2007
Cited by
PubMed Abstract: The green fluorescent protein (GFP) creates its fluorophore by promoting spontaneous peptide backbone cyclization and amino acid oxidation chemistry on its own Ser65, Tyr66, Gly67 tripeptide sequence. Here we use high-resolution crystallography and mutational analyses to characterize GFP variants that undergo backbone cyclization followed by either anticipated chromophore synthesis via Y66F Calpha-Cbeta double-bond formation or unprecedented loss of a Y66F benzyl moiety via Calpha-Cbeta bond cleavage. We discovered a Y66F cleavage variant that subsequently incorporates an oxygen atom, likely from molecular oxygen, at the Y66 Calpha position. The post-translational products identified from these Y66F GFP structures support a common intermediate that partitions between Calpha-Cbeta oxidation and homolytic cleavage pathways. Our data indicate that Glu222 is the branchpoint control for this partitioning step and also influences subsequent oxygen incorporation reactions. From these results, we propose mechanisms for Y66F Calpha-Cbeta cleavage, oxygen incorporation, and chromophore biosynthesis with shared features that include radical chemistry. By revealing how GFP and RFP protein environments steer chemistry to favor fluorophore biosynthesis and disfavor alternative reactivity, we identify strategies for protein design. The proposed, common, one-electron oxidized, radical intermediate for post-translation modifications in the GFP family has general implications for how proteins drive and control spontaneous post-translational chemical modifications in the absence of metal ions.
PubMed: 17326633
DOI: 10.1021/ja063983u
主引用文献が同じPDBエントリー
実験手法
X-RAY DIFFRACTION (1.6 Å)
構造検証レポート
Validation report summary of 2hgd
検証レポート(詳細版)ダウンロードをダウンロード

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件を2024-11-06に公開中

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