Applicability of new spin trap agent, 2-diphenylphosphinoyl-2-methyl-3,4-dihydro-2H-pyrrole N-oxide, in biological system

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• 作者：Tomohiro Karakawa ; Keizo Sato ; Yosuke Muramoto ; Yoshihiro Mitani ; Masataka Kitamado ; Tatsuya Iwanaga ; Tetsuji Nabeshima ; Kumiko Maruyama ; Kazuko Nakagawa ; Kazuhiko Ishida ; Kazumi Sasamoto
• 关键词：2 ; 2-Dimethyl-3 ; 4-dihydro-2H-pyrrole N-oxide (DMPO) ; 2-Diphenylphosphinoyl-2-methyl-3 ; 4-dihydro-2H-pyrrole N-oxide (DPhPMPO) ; Free radical ; Electron spin resonance ; Human neutrophil
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2008
• 期刊代码：159_0006291x
• 类别：bio
• 出版时间：23 May 2008
• 卷：370
• 期：1
• 页码：93-97
• 文件大小：297 K

摘要

Electron spin resonance using spin-trapping is a useful technique for detecting direct reactive oxygen species, such as superoxide (BK-4S32BXM-4&_mathId=mml6&_user=1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=75c9f342e297415cb6c0e4b31740f0ae">). However, the widely used spin trap 2,2-dimethyl-3,4-dihydro-2H-pyrrole N-oxide (DMPO) has several fundamental limitations in terms of half-life and stability. Recently, the new spin trap 2-diphenylphosphinoyl-2-methyl-3,4-dihydro-2H-pyrrole N-oxide (DPhPMPO) was developed by us. We evaluated the biological applicability of DPhPMPO to analyze 1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=377969901c51ef4d55f998c1ae302ad5"> in both cell-free and cellular systems. DPhPMPO had a larger rate constant for 1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=88e082aaf6e4b078cb2b68f430087681"> and formed more stable spin adducts for 1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=506813ef25e60f645a2503284cbb781e"> than DMPO in the xanthine/xanthine oxidase (X/XO) system. In the phorbol myristate acetate-activated neutrophil system, the detection potential of DPhPMPO for 10">10&_user=1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=49bf8861e157bcf3d2b7461f328f8fde"> was significantly higher than that of DMPO (k_DMPO = 13.95 M⁻¹ s⁻¹, k_DPhPMPO = 42.4 M⁻¹ s⁻¹). These results indicated that DPhPMPO is a potentially good candidate for trapping 1067359&_cdi=6713&_rdoc=20&_acct=C000050221&_version=1&_userid=10&md5=39a04ecf23f81892b0c134bc22d0ad38"> in a biological system.