Sensing Caspase 3 Activity with Quantum Dot-Fluorescent Protein Assemblies

被引:249
作者
Boeneman, Kelly [1 ]
Mei, Bing C. [3 ,5 ]
Dennis, Allison M. [4 ]
Bao, Gang [4 ]
Deschamps, Jeffrey R. [2 ]
Mattoussi, Hedi [3 ]
Medintz, Igor L. [1 ]
机构
[1] USN, Res Lab, Ctr Biomol Sci & Engn, Washington, DC 20375 USA
[2] USN, Res Lab, Struct Matter Lab, Washington, DC 20375 USA
[3] USN, Res Lab, Div Opt Sci, Washington, DC 20375 USA
[4] Georgia Inst Technol, Dept Biomed Engn, Atlanta, GA 30332 USA
[5] Univ Massachusetts, Dept Chem Engn, Amherst, MA 01003 USA
关键词
RESONANCE ENERGY-TRANSFER; INHIBITORS; APOPTOSIS; FRET; PEPTIDES; CANCER; PROBES; PAIRS;
D O I
10.1021/ja809721j
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We demonstrate the use of a hybrid fluorescent protein semiconductor quantum dot (QD) sensor capable of specifically monitoring caspase 3 proteolytic activity. mCherry monomeric red fluorescent protein engineered to express an N-terminal caspace 3 cleavage site was ratiometricalty self-assembled to the surface of QDs using metal-affinity coordination. The proximity of the fluorescent protein to the QD allows it to function as an efficient fluorescence resonance energy transfer acceptor. Addition of caspase 3 enzyme to the QD-mCherry conjugates specifically cleaved the engineered mCherry tinker sequence, altering the energy transfer with the QD and allowing quantitative monitoring of proteolytic activity. Inherent advantages of this sensing approach include bacterial. expression of the protease substrate in a fluorescently 1 appended form, facile self-assembly to QDs, and the ability to recombinantly modify the substrate to target other proteases of interest.
引用
收藏
页码:3828 / +
页数:4
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