Determination of rate constants of the reactions of thiols with superoxide radical by electron paramagnetic resonance: Critical remarks on spectrophotometric approaches

被引:61
作者
Dikalov, S
Khramtsov, V
Zimmer, G
机构
[1] UNIV FRANKFURT KLINIKUM,GUSTAV EMBDEN ZENTRUM BIOL CHEM,D-60590 FRANKFURT,GERMANY
[2] INST CHEM KINET & COMBUST,NOVOSIBIRSK,RUSSIA
关键词
D O I
10.1006/abbi.1996.0067
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two new EPR approaches were developed for determination of rate constants of reaction of glutathione (GSH), N-(2-mercaptopropionyl)glycine (MPG), dihydrolipoic acid (DHL), bisnor-dihydrolipoic acid (BNL), and tetranor-dihydrolipoic acid (TNL) with superoxide radical, In both cases the competition between thiols and spin-trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) for superoxide radical was used, In the first method the dependence of amplitude of EPR spectrum of DMPO-OOH spin adduct on concentration of thiols in a superoxide-generating system was studied, In the second approach the changes in reduced thiol concentration due to reaction with superoxide radical were measured by nitroxide biradical containing disulfide bond, Observed rate constants were the following: GSH, 1.8 X 10(5) M(-1) s(-1); MPG, 2.2 X 10(5) M(-1) s(-1); TNL, 1.2 X 10(5) M(-1) s(-1); BNL, 2.5 X 10(5) M(-1) s(-1); DHL, 4.8 X 10(5) M(-1) s(-1). The determination of the rate constants of reaction of superoxide radical with thiols by spectrophotometrical cytochrome C assay could result in an underestimation of the values due to the reduction of cytochrome C by thiols. Use of epinephrine for this purpose could lead to an overestimation of experimental rate constants because the adrenochrome formed in the reaction of epinephrine with superoxide radical reacts with thiols. (C) 1996 Academic Press, Inc.
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页码:207 / 218
页数:12
相关论文
共 38 条
  • [1] ASMUS KD, 1990, METHOD ENZYMOL, V186, P168
  • [2] BACHUR NR, 1977, MOL PHARMACOL, V38, P1745
  • [3] REACTIVITY OF HO2/O-2 RADICALS IN AQUEOUS-SOLUTION
    BIELSKI, BHJ
    CABELLI, DE
    ARUDI, RL
    ROSS, AB
    [J]. JOURNAL OF PHYSICAL AND CHEMICAL REFERENCE DATA, 1985, 14 (04) : 1041 - 1100
  • [4] MARKED REDUCTION OF FREE-RADICAL GENERATION AND CONTRACTILE DYSFUNCTION BY ANTIOXIDANT THERAPY BEGUN AT THE TIME OF REPERFUSION - EVIDENCE THAT MYOCARDIAL STUNNING IS A MANIFESTATION OF REPERFUSION INJURY
    BOLLI, R
    JEROUDI, MO
    PATEL, BS
    ARUOMA, OI
    HALLIWELL, B
    LAI, EK
    MCCAY, PB
    [J]. CIRCULATION RESEARCH, 1989, 65 (03) : 607 - 622
  • [5] REMOVAL OF FERRITIN-BOUND IRON BY DL-DIHYDROLIPOATE AND DL-DIHYDROLIPOAMIDE
    BONOMI, F
    PAGANI, S
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1986, 155 (02): : 295 - 300
  • [6] KINETIC APPLICATIONS OF ELECTRON PARAMAGNETIC RESONANCE SPECTROSCOPY .3. SELF-REACTIONS OF DIALKYL NITROXIDE RADICALS
    BOWMAN, DF
    GILLAN, T
    INGOLD, KU
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1971, 93 (24) : 6555 - &
  • [7] BUETTNER GR, 1990, METHOD ENZYMOL, V186, P127
  • [8] CADENAS E, 1990, METHOD ENZYMOL, V186, P180
  • [9] ROLE OF IRON-ION CHELATION BY QUINONES IN THEIR REDUCTION, OH-RADICAL GENERATION AND LIPID-PEROXIDATION
    DIKALOV, S
    ALOV, P
    RANGELOVA, D
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1993, 195 (01) : 113 - 119
  • [10] 2H-IMIDAZOLE-N-OXIDES AS SPIN TRAPS
    DIKALOV, SI
    KIRILYUK, IA
    GRIGOREV, IA
    VOLODARSKII, LB
    [J]. BULLETIN OF THE RUSSIAN ACADEMY OF SCIENCES-DIVISION OF CHEMICAL SCIENCE, 1992, 41 (05): : 834 - 837