1RMP

Probing the Role of Tryptophans in Aequorea Victoria Green Fluorescent Proteins with an Expanded Genetic Code

1RMP の概要

• エントリーDOI: 10.2210/pdb1rmp/pdb
• 関連するPDBエントリー: 1EMG 1RM9 1RMM 1RMO
• 分子名称: SIGF1-GFP fusion protein (2 entities in total)
• 機能のキーワード: beta-barrel; gfp; noncanonical amino acid, 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: 25726.82

構造登録者

Budisa, N.,Pal, P.P.,Alefelder, S.,Birle, P.,Krywcun, T.,Rubini, M.,Wenger, W.,Bae, J.H.,Steiner, T. (登録日: 2003-11-28, 公開日: 2004-06-08, 最終更新日: 2023-11-15)

主引用文献

Budisa, N.,Pal, P.P.,Alefelder, S.,Birle, P.,Krywcun, T.,Rubini, M.,Wenger, W.,Bae, J.H.,Steiner, T.
Probing the role of tryptophans in Aequorea victoria green fluorescent proteins with an expanded genetic code
Biol.Chem., 385:191-202, 2004

PubMed Abstract: The expanded genetic code in combination with site-directed mutagenesis was used to probe spectroscopic and structural roles of tryptophan (Trp) residues in Aequorea victoria green fluorescent proteins (avGFPs). Nine different halogen-, chalcogen-, and methyl-containing Trp isosteric analogues and surrogates were incorporated into avGFPs containing indole moieties in, and outside of, the chromophore, by the use of the selective pressure incorporation method. Such isosteric replacements introduced minimal local geometry changes in indole moieties, often to the level of single atomic exchange ('atomic mutation') and do not affect three-dimensional structures of avGFPs but induce changes in spectral properties. Our approach offers a new platform to re-evaluate issues like resonance transfer, mechanisms of chromophore formation and maturation, as well as the importance of local geometry and weak sulphur-aromatic interactions for avGFP spectral properties and structural stability. The library of novel tailor-made avGFP mutants and variants generated in this work has demonstrated not only the potentials of the expanded genetic code to study spectroscopic functions, but also a new approach to generate tailor-made proteins with interesting and useful spectral properties.

PubMed: 15101562
DOI: 10.1515/BC.2004.038

実験手法

X-RAY DIFFRACTION (3 Å)