EPR METHOD FOR THE MEASUREMENT OF CELLULAR SULFHYDRYL-GROUPS

被引:29
作者
WEINER, LM
HU, H
SWARTZ, HM
机构
[1] UNIV ILLINOIS,COLL MED,URBANA,IL 61801
[2] EPR RES CTR,URBANA,IL 61801
关键词
NITROXYL BIRADICAL; THIOL-DISULFIDE EXCHANGE; EPR; INTACT CELL; GLUTATHIONE CONTENT;
D O I
10.1016/0014-5793(91)81270-I
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
An EPR method that can measure the concentration of sulfhydryl groups in intact cells has been developed using a specially designed stable nitroxyl biradical. The biradical, RS-SR, contains a disulfide bond and readily undergoes thiol-disulide exchange reactions with thiols resulting in a characteristic EPR spectrum which can be analyzed to provide a quantitative measure of sulfhydryl groups. The data obtained from the EPR method are in good agreement with those obtained from the conventional optical method using Ellman's reagent. The advantages of the EPR method are that the measurement can be carried out on intact cells or any other highly colored, absorbing and/or scattering solutions and the sensitivity is such that only a few cells (approximately 100) are needed for each quantitative measurement.
引用
收藏
页码:243 / 246
页数:4
相关论文
共 14 条
[1]   METHODOLOGY FOR ANALYSIS OF TISSUE SULFHYDRYL COMPONENTS [J].
BOYNE, AF ;
ELLMAN, GL .
ANALYTICAL BIOCHEMISTRY, 1972, 46 (02) :639-&
[2]   KINETICS OF ENZYME-MEDIATED REDUCTION OF LIPID SOLUBLE NITROXIDE SPIN LABELS BY LIVING CELLS [J].
CHEN, K ;
MORSE, PD ;
SWARTZ, HM .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 943 (03) :477-484
[3]  
Cohen G, 1988, PROGR PARKINSON RES, P119
[4]   NOTE ON MOLAR ABSORPTIVITY OF REDUCED ELLMANS REAGENT, 3-CARBOXYLATO-4-NITROTHIOPHENOLATE [J].
COLLIER, HB .
ANALYTICAL BIOCHEMISTRY, 1973, 56 (01) :310-311
[5]  
DOLPHIN A, 1989, GLUTATIONE CHEM BIOC
[6]  
JAKOBY WB, 1987, METHOD ENZYMOL, V143
[7]   NITROXIDES AS POTENTIAL CONTRAST ENHANCING AGENTS FOR MRI APPLICATION - INFLUENCE OF STRUCTURE ON THE RATE OF REDUCTION BY RAT HEPATOCYTES, WHOLE LIVER HOMOGENATE, SUBCELLULAR-FRACTIONS, AND ASCORBATE [J].
KEANA, JFW ;
POU, S ;
ROSEN, GM .
MAGNETIC RESONANCE IN MEDICINE, 1987, 5 (06) :525-536
[8]   QUANTITATIVE-DETERMINATION OF SH-GROUPS IN LOW-MOLECULAR-WEIGHT AND HIGH-MOLECULAR-WEIGHT COMPOUNDS BY AN ELECTRON-SPIN RESONANCE METHOD [J].
KHRAMTSOV, VV ;
YELINOVA, VI ;
WEINER, LM ;
BEREZINA, TA ;
MARTIN, VV ;
VOLODARSKY, LB .
ANALYTICAL BIOCHEMISTRY, 1989, 182 (01) :58-63
[9]   GLUTATHIONE [J].
MEISTER, A ;
ANDERSON, ME .
ANNUAL REVIEW OF BIOCHEMISTRY, 1983, 52 :711-760
[10]  
METZLER DE, 1977, BIOCHEMISTRY-US, pCH7