Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors

被引：63

作者：

Bermejo, Clara ^{[1
]}

Haerizadeh, Farzad ^{[1
]}

Takanaga, Hitomi ^{[1
]}

Chermak, Diane ^{[1
]}

Frommer, Wolf B. ^{[1
]}

机构：

[1] Carnegie Inst Sci, Dept Plant Biol, Stanford, CA 94305 USA

来源：

BIOCHEMICAL JOURNAL | 2010年 / 432卷

基金：

美国国家卫生研究院;

关键词：

Forster resonance energy transfer (FRET); hexose transport; metabolic flux; microfluidic platform; starvation; SACCHAROMYCES-CEREVISIAE; HEXOSE TRANSPORTERS; HXT5; EXPRESSION; CELLS; VISUALIZATION; REVEALS; PROTEIN; FAMILY; GENES;

D O I：

10.1042/BJ20100946

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：

Precise and dynamic measurement of intracellular metabolite levels has been hampered by difficulties in differentiating between adsorbed and imported fractions and the subcellular distribution between cytosol, endomembrane compartments and mitochondria. In the present study, genetically encoded FRET (Forster resonance energy transfer)-based sensors were deployed for dynamic measurements of free cytosolic glucose and ATP with varying external supply and in glucose-transport mutants. Moreover, by using the FRET sensors in a microfluidic platform, we were able to monitor in vivo changes of intracellular free glucose in individual yeast cells. We demonstrate the suitability of the FRET sensors for gaining physiological insight by demonstrating that free intracellular glucose and ATP levels are reduced in a hxt5 Delta hexose-transporter mutant compared with wild-type and other hxt Delta strains.

引用

页码：399 / 406

页数：8

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