68498-25-9Relevant articles and documents
A New One-Pot Fluorescence Derivatization Strategy for Highly Sensitive MicroRNA Analysis
Pan, Li,Zhang, Huaisheng,Zhao, Jingjin,Ogungbe, Ifedayo Victor,Zhao, Shulin,Liu, Yi-Ming
, p. 5639 - 5647 (2020/03/23)
MicroRNAs (miRNAs) modulate the expression of over 30 % of mammalian genes during development and apoptosis, and abnormal expression of miRNAs may lead to a range of human pathologies. Therefore, analysis of miRNAs is valuable for disease diagnostics. In this work, a novel one-pot fluorescence derivatization strategy was developed for miRNA analysis. The mechanism of the derivatization reaction was explored by using instrumental methods, including liquid chromatography, fluorescence spectroscopy, and mass spectrometry. Highly fluorescent N6-ethenoadenine (?-adenine) was formed and detached from the miRNA sequence through the reaction of adenine in nucleic acids with 2-chloroacetaldehyde (CAA) at 100 °C. This is the first experimental evidence that the cooperation of formed ?-adenine and water-mediated hydrogen-bond interaction between the proton at the 2′- and the oxyanion at 3′-positions stabilized the oxocarbenium significantly, which makes the depurination and derivatization of miRNA highly effective. Based on this derivatization strategy, a facile and sensitive high-performance liquid chromatography method was developed for quantitative assay of miRNAs. In combination with magnetic solid-phase extraction (MSPE), the HPLC method was shown to be useful for the determination of microRNAs at sub-picomolar level in serum samples.
In vitro synthesis of 1,N6-etheno-2′-deoxyadenosine and 1,N2-etheno-2′-deoxyguanosine by 2,4-dinitrophenol and 1,3-dinitropyrene in presence of a bacterial nitroreductase
Chiron, Serge,Barbati, Stephane,De Meo, Michel,Botta, Alain
, p. 222 - 227 (2007/10/03)
The formation of covalent nitro-PAH DNA adducts and nitro-PAH mediated oxidative lesions are two possible mechanisms for the initiation of nitro-PAH carcinogenesis. Sixty-minute incubation of 1,3-dinitropyrene (100 μM) or 1,4-dinitrophenol (100 μM) with a mixture of 150 μM NADH, 0.5 units of E. coli nitroreductase, 100 μM linoleic acid, 0.5 mM ferrous iron, and 100 μM 2′-deoxyadenosine (2′-dA) or 100 μM 2′-deoxyguanosine (2′-dG) were analyzed by liquid chromatography multistage mass spectrometry. Mixtures of 1,N6-etheno-2′-deoxyadenosine (εdA) plus 4-oxo-2-nonenal (4-ONE) and 1,N2-etheno-2′- deoxyguanosine (εdG) plus 4-ONE could be detected from 2′-dA and 2′-dG, respectively. Addition of 2% propanol inhibited the formation of etheno adducts. Analyses of disappearance kinetics of dA and dG showed that dG was more rapidly eliminated than does dA (t[1/2] = 23.3 min and 98.3 min for dG and dA, respectively). Curves of formation kinetics revealed that the peak of εdG was at 55.6 min while that of εdA was at 186.9 min. These peaks represented 1.43% and 1.25% of the original dG and dA, respectively. In both cases, the peaks were followed by rapid degradations of etheno adducts. The results, obtained in this system, do not allow any extrapolation to realistic cellular responses; nevertheless, these data questioned the validity of the use of unsubstituted etheno adducts as reliable oxidative stress and nitro-PAH exposure biomarkers.
4,5-Epoxy-2(E)-decenal-induced formation of 1,N6-etheno-2′-deoxyadenosine and 1,N2-etheno-2′-deoxyguanosine adducts
Lee, Seon Hwa,Oe, Tomoyuki,Blair, Ian A.
, p. 300 - 304 (2007/10/03)
Trans-4,5-Epoxy-2(E)-decenal reacted with 2′-deoxyadenosine to give 1,N6-etheno-2′-deoxyadenosine as well as other 2′-deoxyadenosine adducts. It also reacted with 2′-deoxyguanosine to give 1,N2-etheno-2′-deoxyguanosine and other 2′-deoxyguanosine adducts. Synthetic trans-4,5-epoxy-2(E)-decenal was quite stable under the reaction conditions that were used. It was not contaminated with 2,3-epoxyoctanal, a potential precursor to the formation of unsubstituted etheno adducts. Furthermore, using a sensitive LC/MS assay, it was possible to show that no 2,3-epoxyoctanal was formed during prolonged incubations of trans-4,5-epoxy-2(E)-decenal. Therefore, trans-4,5-epoxy-2(E)-decenal, a primary product of lipid peroxidation, is a precursor to the formation of 1,N6-etheno-2′-deoxyadenosine and 1,N2-etheno-2′-deoxyguanosine. There is no need for an additional oxidation step such as would be required if trans,trans-2,4-decadienal or 4-hydroxy-2-nonenal were the lipid hydroperoxide decomposition products that initiated the formation of unsubstituted etheno adducts. These findings provide an important link between a primary product of lipid peroxidation and a mutagenic DNA lesion that has been detected in human tissues.
