Environmentally Robust Rhodamine Reporters for Probe-based Cellular Detection of the Cancer-linked Oxidoreductase hNQO1
| Autor: | Robin L. McCarley, Quinn A. Best, Amanda E. Johnson, Bijeta Prasai, Alexandra Rouillere |
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| Rok vydání: | 2015 |
| Předmět: |
010402 general chemistry
Photochemistry 01 natural sciences Biochemistry Rhodamines Rhodamine chemistry.chemical_compound Oxidoreductase Cell Line Tumor Neoplasms NAD(P)H Dehydrogenase (Quinone) Humans Fluorescent Dyes Xanthene chemistry.chemical_classification Molecular Structure 010405 organic chemistry General Medicine Fluorescence 0104 chemical sciences Kinetics chemistry Molecular Medicine Chemical stability NAD+ kinase |
| Zdroj: | ACS Chemical Biology. 11:231-240 |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/acschembio.5b00792 |
| Popis: | We successfully synthesized a fluorescent probe capable of detecting the cancer-associatedquinoneoxidoreductase isozyme-1 within human cells, based on results from an investigation of the stability of various rhodamines and seminaphthorhodamines toward the biological reductant NADH, present at ∼100-200 μM within cells. While rhodamines are generally known for their chemical stability, we observe that NADH causes significant and sometimes rapid modification of numerous rhodamine analogues, including those oftentimes used in imaging applications. Results from mechanistic studies lead us to rule out a radical-based reduction pathway, suggesting rhodamine reduction by NADH proceeds by a hydride transfer process to yield the reduced leuco form of the rhodamine and oxidized NAD(+). A relationship between the structural features of the rhodamines and their reactivity with NADH is observed. Rhodamines with increased alkylation on the N3- and N6-nitrogens, as well as the xanthene core, react the least with NADH; whereas, nonalkylated variants or analogues with electron-withdrawing substituents have the fastest rates of reaction. These outcomes allowed us to judiciously construct a seminaphthorhodamine-based, turn-on fluorescent probe that is capable of selectively detecting the cancer-associated, NADH-dependent enzymequinoneoxidoreductase isozyme-1 in human cancer cells, without the issue of NADH-induced deactivation of the seminaphthorhodamine reporter. |
| Databáze: | OpenAIRE |
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