Synthesis and Tritium Labeling of New Aromatic Diazirine Building Blocks for Photoaffinity Labeling and Cross-Linking

Ambroise, Yves; Pillon, Florence; Mioskowski, Charles; Valleix, Alain; Rousseau, Bernard

doi:10.1002/1099-0690(200110)2001:20<3961::aid-ejoc3961>3.0.co;2-r

Cited by 24 publications

(11 citation statements)

References 18 publications

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“…2). The diazirine group of 2 showed a characteristic absorbance peak at about 348 nm, consistent with the spectral properties of other reported 3-trifluoromethyl-3-aryldiazirines 26-29. Upon UV irradiation, the absorbance at 348 nm decreased with a half-life of about 40 s following approximately first-order kinetics (Fig.…”

Section: Resultssupporting

confidence: 87%

A diazirine-based photoaffinity etoposide probe for labeling topoisomerase II

Chee

Yalowich

Bodner

et al. 2010

Bioorganic & Medicinal Chemistry

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Etoposide is a widely used anticancer drug that targets topoisomerase II, an essential nuclear enzyme. However, despite the fact that it has been in use and studied for more than 30 years the specific site on the enzyme to which it binds is unknown. In order to identify the etoposide binding site(s) on topoisomerase II, a diazirine-based photoaffinity etoposide analog probe has been synthesized and its photoreactivity and biological activities have been characterized. Upon UV irradiation, the diazirine probe rapidly produced a highly reactive carbene species that formed covalent adducts containing stable carbon-based bonds indicating that it should also be able to form stable covalent adducts with amino acid residues on topoisomerase II. The human leukemia K562 cell growth and topoisomerase II inhibitory properties of the diazirine probe suggest that it targets topoisomerase II in a manner similar to etoposide. The diazirine probe was also shown to act as a topoisomerase II poison through its ability to cause topoisomerase IIα-mediated double-strand cleavage of DNA. Additionally, the diazirine probe significantly increased protein-DNA covalent complex formation upon photoirradiation of diazirine probe-treated K562 cells, as compared to etoposide-treated cells. This result suggests that the photoactivated probe forms a covalent adduct with topoisomerase IIα. In conclusion, the present characterization of the chemical, biochemical, and biological properties of the newly synthesized diazirine-based photoaffinity etoposide analog indicates that use of a proteomics mass spectrometry approach will be a tractable strategy for future identification of the etoposide binding site(s) on topoisomerase II through covalent labeling of amino acid residues.

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Section: Resultssupporting

confidence: 87%

A diazirine-based photoaffinity etoposide probe for labeling topoisomerase II

Chee

Yalowich

Bodner

et al. 2010

Bioorganic & Medicinal Chemistry

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“…The photochemical reactions of chemical probe 3 were examined to confirm its reactivity under photoactivation. As shown in other reports, 3-trifluoromethyl-3-aryldiazirine has a characteristic absorbance peak at about 348 nm [ 23 , 24 , 25 ]. The photochemical kinetics of compound 3 in methanol at excitation wavelength 348 nm for fixed periods of time were measured spectrophotometrically ( Supporting Information Figure 1 ) Since UV absorption of the dansyl group and benzene group of compound 3 is stronger than that of diazirine compound, there was no significant change in absorption spectra.…”

Section: Resultsmentioning

confidence: 55%

Synthesis of a Dual-Labeled Probe of Dimethyl Lithospermate B with Photochemical and Fluorescent Properties

2011

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Dimethyl lithosermate B (DLB) is a highly potent natural antioxidant and antidiabetic polyphenol with unknown mode of action. To determine its cellular targets, a photochemical and fluorescent dimethyl lithopermate B probe was designed and efficiently synthesized. The dual-labeled chemical probe for biological application was evaluated by UV and fluorescence to determine its electrochemical absorption and emission properties. This probe could be valuable for investigating ligand-protein interactions and subcellular localization.

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“…Fipronil ( 25 ) is a significant insecticide acting as a noncompetitive antagonist at the GABA A chloride ion channel 102. Casida designed a tritiated photoaffinity diazirine analogue ( 26 ) of the parent substance,103 and employed a diiodo precursor to efficiently prepare 26 using the chemoselective catalytic tritium deiodination strategy (Scheme )104 reported by Rousseau 105–107. The final example of the work in this area by the Casida group is perhaps the most intriguing story from an artistic, cultural and historical perspective.…”

Section: Gaba Receptor Antagonistsmentioning

confidence: 99%