FEBS Letters | |
Detection of superoxide anion using an isotopically labeled nitrone spin trap: potential biological applications | |
Joseph, Joy1  Kalyanaraman, B1  Vasquez-Vivar, Jeannette4  Nsanzumuhire, Cline3  Martásek, Pavel2  Zhang, Hao1  Tordo, Paul3  Karoui, Hakim3  | |
[1] Biophysics Research Institute, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA;University of Texas Health Sciences Center, San Antonio, TX, USA;Laboratoire ‘Structure et Réactivité des Espèces Paramagnétiques’, Université de Provence, Marseille, France;Department of Pathology and Cardiovascular Research Center, Medical College of Wisconsin, Milwaukee, WI, USA | |
关键词: Electron spin resonance; Spin trap; Nitric oxide synthase; Vascular biology; Tetrahydrobiopterin; ESR; electron spin resonance; DMPO; 5; 5-dimethyl-1-pyrroline N-oxide; DEPMPO; 5-(dimethoxyphosphoryl)-5′-methyl-1-pyrroline N-oxide; EMPO; 5-ethoxycarbonyl-5-methyl-pyrroline N-oxide; nNOS; neuronal nitric oxide synthase; eNOS; endothelial nitric oxide synthase; DMPO–OOH; DMPO superoxide adduct; DEPMPO–OOH; DEPMPO superoxide adduct; EMPO–OOH; EMPO superoxide adduct; EMPO–OH; EMPO hydroxyl adduct; | |
DOI : 10.1016/S0014-5793(00)01498-8 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
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【 摘 要 】
We describe the synthesis and biological applications of a novel nitrogen-15-labeled nitrone spin trap, 5-ethoxycarbonyl-5-methyl-1-pyrroline N-oxide ([15N]EMPO) for detecting superoxide anion. Superoxide anion generated in xanthine/xanthine oxidase (100 nM min−1) and NADPH/calcium-calmodulin/nitric oxide synthase systems was readily detected using EMPO, a nitrone analog of 5,5′-dimethyl-1-pyrroline N-oxide (DMPO). Unlike DMPO-superoxide adduct (DMPO–OOH), the superoxide adduct of EMPO (EMPO–OOH) does not spontaneously decay to the corresponding hydroxyl adduct, making spectral interpretation less confounding. Although the superoxide adduct of 5-(diethoxyphosphoryl)-5-methyl-pyrroline N-oxide is more persistent than EMPO–OOH, the electron spin resonance spectra of [14N]EMPO–OOH and [15N]EMPO–OOH are less complex and easier to interpret. Potential uses of [15N]EMPO in elucidating the mechanism of superoxide formation from nitric oxide synthases, and in ischemia/reperfusion injury are discussed.
【 授权许可】
Unknown
【 预 览 】
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RO201912020309317ZK.pdf | 195KB | ![]() |