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Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
Published on: May 29, 2011
A high-performance fluorescent probe for detection of cysteine in plasma constructed by combining Cu(I) and
Xiaozong Dou1, Zhenhong Jia2, Hongyan Zhang1
1Xinjiang Key Laboratory of Solid State Physics and Devices, Xinjiang University, Urumqi 830017, China; School of Physical Science and Technology, Xinjiang University, Urumqi 830017, China.
Abstract:
A high-performance fluorescent probe 2,5-dimercapto-1,3,4-thiadiazole copper nanoparticles (DMTD-CuNPs) was synthesized by hydrothermal method based on monovalent copper (Cu(I)) and 2,5-dimercapto-1,3,4-thiadiazole (DMTD), and it can effectively detect cysteine (Cys) in plasma. Experiments show that DMTD can reduces band gap of Cu(I) in DMTD-CuNPs, promote charge transfer transition from DMTD to Cu(I) and significantly enhance fluorescence intensity of DMTD-CuNPs at 515 nm. The large Stokes shift of DMTD-CuNPs is 315 nm, which can reduce the self-quenching of probe fluorescence and improves detection accuracy of the probe. In the presence of Cys, fluorescence of DMTD-CuNPs at 515 nm is significantly quenched because Cys reacts with Cu(I) in DMTD-CuNPs through Cu-S bond to form reduced charge transfer, which can be successfully used for the detection of Cys. Linear range and detection limit for Cys detection are 25-65 µM and 50 nM, respectively. Furthermore, feasibility of detecting Cys in plasma using DMTD-CuNPs probe was evaluated by standard addition method, and the absolute recovery is 96-99%. Such a DMTD-CuNPs probe shows high sensitivity, good selectivity and low detection limit for Cys, which is expected to be used for the practical analysis of Cys in plasma.

