Development of a fluorescent nanosensor for ribose

被引：101
作者：
Lager, I
Fehr, M
Frommer, WB
Lalonde, SW
机构：
[1] ZMBP Tubingen, Plant Physiol, D-72076 Tubingen, Germany
[2] Carnegie Inst, Stanford, CA 94305 USA
来源：
FEBS LETTERS | 2003年 / 553卷 / 1-2期
关键词：
nanosensor; ribose; fluorescence resonance energy transfer; transport;
D O I：
10.1016/S0014-5793(03)00976-1
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：
To analyze ribose uptake and metabolism in living cells, nanosensors were engineered by flanking the Escherichia coli periplasmic ribose binding protein with two green fluorescent protein variants. Following binding of ribose, fluorescence resonance energy transfer decreased with increasing ribose concentration. Five affinity mutants were generated covering binding constants between 400 nM and 11.7 mM. Analysis of nanosensor response in COS-7 cells showed that free ribose accumulates in the cell and is slowly metabolized. Inhibitor studies suggest that uptake is mediated by a monosaccharide transporter of the GLUT family, however, ribose taken up into the cell was not or only slowly released, indicating irreversibility of uptake. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码：85 / 89
页数：5
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