Structural basis for the phototoxicity of the fluorescent protein KillerRed

被引：93

作者：

Carpentier, Philippe ^{[1
]}

Violot, Sebastien ^{[2
]}

Blanchoin, Laurent ^{[2
]}

Bourgeois, Dominique ^{[1
,3
]}

机构：

[1] Univ Grenoble 1, CNRS, CEA, Inst Biol Struct Jean Pierre Ebel,IBS,Lab Cristal, F-38027 Grenoble, France

[2] Univ Grenoble 1, CNRS, INRA, Lab Physiol Cellulaire Vegetale,CEA,Inst Rech Tec, F-38054 Grenoble, France

[3] ESRF, F-38043 Grenoble, France

来源：

FEBS LETTERS | 2009年 / 583卷 / 17期

关键词：

CALI; Fluorescent proteins; X-ray crystallography; Reactive oxygen species; Solvent channel; ASSISTED LASER INACTIVATION; EGFP-FUSION PROTEINS; LIGHT INACTIVATION; CHROMOPHORE; CELLS; DECARBOXYLATION; PHOTOSENSITIZER; CORAL; CALI;

D O I：

10.1016/j.febslet.2009.07.041

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The red fluorescent protein KillerRed, engineered from the hydrozoan chromoprotein anm2CP, has been reported to induce strong cytotoxicity through the chromophore assisted light inactivation (CALI) effect. Here, we present the X-ray structures of KillerRed in its native and bleached states. A long water-filled channel is revealed, connecting the methylene bridge of the chromophore to the solvent. This channel facilitates the transit of oxygen and of reactive oxygen species (ROS) formed by reaction with the excited chromophore. The functional roles of key mutations used to produce KillerRed are discussed, strong chromophore distortions in the bleached state are revealed, and mechanisms for ROS production and self protection are proposed. The presence of a partially mature, photo-resistant, green-emitting state is characterized, which accounts for enhanced CALI by "pre-bleached" KillerRed. (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.

引用

页码：2839 / 2842

页数：4

共 24 条

[21] Enhanced EGFP-chromophore-assisted laser inactivation using deficient cells rescued with functional EGFP-fusion proteins [J].