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Biosens Bioelectron. 2016 Nov 15;85:437-444. doi: 10.1016/j.bios.2016.04.101. Epub 2016 May 13.

Development of new peptide-based receptor of fluorescent probe with femtomolar affinity for Cu(+) and detection of Cu(+) in Golgi apparatus.

Author information

1
Bioorganic Chemistry Laboratory, Center for Design and Applications of Molecular Catalysts, Department of Chemistry and Chemical Engineering, Inha University, Incheon 402-751, South Korea.
2
Department of Biomedical Sciences, Inha University College of Medicine, Incheon 402-751, South Korea; Hypoxia-related Disease Research Center, College of Medicine, Inha University, Incheon 402-751, South Korea.
3
Hypoxia-related Disease Research Center, College of Medicine, Inha University, Incheon 402-751, South Korea; Department of Microbiology, Inha Research Institute for Medical Science, College of Medicine, Inha University, Incheon 402-751, South Korea. Electronic address: park001@inha.ac.kr.
4
Bioorganic Chemistry Laboratory, Center for Design and Applications of Molecular Catalysts, Department of Chemistry and Chemical Engineering, Inha University, Incheon 402-751, South Korea. Electronic address: leekh@inha.ac.kr.

Abstract

Developing fluorescent probes for monitoring intracellular Cu(+) is important for human health and disease, whereas a few types of their receptors showing a limited range of binding affinities for Cu(+) have been reported. In the present study, we first report a novel peptide receptor of a fluorescent probe for the detection of Cu(+). Dansyl-labeled tripeptide probe (Dns-LLC) formed a 1:1 complex with Cu(+) and showed a turn-on fluorescent response to Cu(+) in aqueous buffered solutions. The dissociation constant of Dns-LLC for Cu(+) was determined to be 12 fM, showing that Dns-LLC had more potent binding affinity for Cu(+) than those of previously reported chemical probes for Cu(+). The binding mode study showed that the thiol group of the peptide receptor plays a critical role in potent binding with Cu(+) and the sulfonamide and amide groups of the probe might cooperate to form a complex with Cu(+). Dns-LLC detected Cu(+) selectively by a turn-on response among various biologically relevant metal ions, including Cu(2+) and Zn(2+). The selectivity of the peptide-based probe for Cu(+) was strongly dependent on the position of the cysteine residue in the peptide receptor part. The fluorescent peptide-based probe penetrated the living RKO cells and successfully detected Cu(+) in the Golgi apparatus in live cells by a turn-on response. Given the growing interest in imaging Cu(+) in live cells, a novel peptide receptor of Cu(+) will offer the potential for developing a variety of fluorescent probes for Cu(+) in the field of copper biochemistry.

KEYWORDS:

Chemosensor; Copper; Cu(+); Fluorescent; Peptide; Probe

PMID:
27208475
DOI:
10.1016/j.bios.2016.04.101
[Indexed for MEDLINE]

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