HyPer-3: A Genetically Encoded H2O2 Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging

被引：354

作者：

Bilan, Dmitry S. ^{[1
,2
]}

Pase, Luke ^{[4
]}

Joosen, Linda ^{[5
,6
]}

Gorokhovatsky, Andrey Yu ^{[2
,7
]}

Ermakova, Yulia G. ^{[2
,3
]}

Gadella, Theodorus W. J. ^{[5
,6
]}

Grabher, Clemens ^{[4
]}

Schultz, Carsten ^{[8
]}

Lukyanov, Sergey ^{[1
,2
]}

Belousov, Vsevolod V. ^{[1
,2
]}

机构：

[1] Nizhny Novgorod State Med Acad, Nizhnii Novgorod, Russia

[2] Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia

[3] Moscow MV Lomonosov State Univ, Fac Biol, Moscow, Russia

[4] Forschungszentrum Karlsruhe, Inst Toxicol & Genet, Karlsruhe Inst Technol, D-76344 Eggenstein Leopoldshafen, Germany

[5] Univ Amsterdam, Swammerdam Inst Life Sci, NL-1098 XH Amsterdam, Netherlands

[6] Univ Amsterdam, Netherlands Inst Syst Biol, NL-1098 XH Amsterdam, Netherlands

[7] Shemyakin Ovchinnikov Inst Bioorgan Chem, Pushchino Branch, Pushchino, Russia

[8] European Mol Biol Lab, D-69117 Heidelberg, Germany

来源：

ACS CHEMICAL BIOLOGY | 2013年 / 8卷 / 03期

基金：

俄罗斯基础研究基金会;

关键词：

TRANSCRIPTIONAL REGULATOR OXYR; HYDROGEN-PEROXIDE; REVERSIBLE INACTIVATION; MUTATIONAL ANALYSIS; REGIONS IMPORTANT; IDENTIFICATION; MICROSCOPY; FLIM; ROS;

D O I：

10.1021/cb300625g

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

High-performance sensors for reactive oxygen species are instrumental to monitor dynamic events in cells and organisms. Here, we present HyPer-3, a genetically encoded fluorescent indicator for intracellular H2O2 exhibiting improved performance with respect to response time and speed. HyPer-3 has an expanded dynamic range compared to HyPer and significantly faster oxidation/reduction dynamics compared to HyPer-2. We demonstrate this performance by in vivo imaging of tissue scale H2O2 gradients in zebrafish larvae. Moreover, HyPer-3 was successfully employed for single wavelength fluorescent lifetime imaging of H2O2 levels both in vitro and in vivo.

引用

页码：535 / 542

页数：8

共 36 条

[1]

Regulation of the OxyR transcription factor by hydrogen peroxide and the cellular thiol -: disulfide status [J].

Åslund, F ;

Zheng, M ;

Beckwith, J ;

Storz, G .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (11) :6161-6165

[2]

Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell [J].

Bastiaens, PIH ;

Squire, A .

TRENDS IN CELL BIOLOGY, 1999, 9 (02) :48-52

[3]

The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology [J].

Bedard, Karen ;

Krause, Karl-Heinz .

PHYSIOLOGICAL REVIEWS, 2007, 87 (01) :245-313

[4]

Genetically encoded fluorescent indicator for intracellular hydrogen peroxide [J].

Belousov, VV ;

Fradkov, AF ;

Lukyanov, KA ;

Staroverov, DB ;

Shakhbazov, KS ;

Terskikh, AV ;

Lukyanov, S .

NATURE METHODS, 2006, 3 (04) :281-286

[5]

Berg J, 2009, NAT METHODS, V6, P161, DOI [10.1038/NMETH.1288, 10.1038/nmeth.1288]

[6]

Structural basis of the redox switch in the OxyR transcription factor [J].

Choi, HJ ;

Kim, SJ ;

Mukhopadhyay, P ;

Cho, S ;

Woo, JR ;

Storz, G ;

Ryu, SE .

CELL, 2001, 105 (01) :103-113

[7]

ROS as signalling molecules:: mechanisms that generate specificity in ROS homeostasis [J].

D'Autreaux, Benoit ;

Toledano, Michel B. .

NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2007, 8 (10) :813-824

[8]

Specific and reversible inactivation of protein tyrosine phosphatases by hydrogen peroxide: Evidence for a sulfenic acid intermediate and implications for redox regulation [J].

Denu, JM ;

Tanner, KG .

BIOCHEMISTRY, 1998, 37 (16) :5633-5642

[9]

Dixon M., 1979, Enzymes.

[10]

Free radicals in the physiological control of cell function [J].

Dröge, W .

PHYSIOLOGICAL REVIEWS, 2002, 82 (01) :47-95

← 1 2 3 4 →