CHARACTERIZATION OF THE IRON-SULFUR CENTERS IN SUCCINATE-DEHYDROGENASE

被引:31
作者
COLES, CJ
HOLM, RH
KURTZ, DM
ORMEJOHNSON, WH
RAWLINGS, J
SINGER, TP
WONG, GB
机构
[1] VET ADM HOSP,DIV MOLEC BIOL,SAN FRANCISCO,CA 94121
[2] STANFORD UNIV,DEPT CHEM,STANFORD,CA 94305
[3] UNIV WISCONSIN,DEPT BIOCHEM,MADISON,WI 53706
关键词
D O I
10.1073/pnas.76.8.3805
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Two techniques have been applied to the determination of the number and type (2-Fe, 4-Fe) of iron-sulfur centers in the iron-sulfur flavoprotein succinate dehydrogenase [succinate:(acceptor) oxidoreductase, EC 1.3.99.1]. One procedure uses p-CF3C6H4SH as an extrusion reagent and Fourier transform 19F nuclear magentic resonance as the method of detection and quantitation of extruded cores of these centers in the form of [Fe2S2(SRF)4]2- and [Fe4S4(SRF)4]2- (RF = p-C6H4CF3). The second procedure, interprotein core transfer, involves thiol displacement of iron-sulfur cores followed by specific core transfer to the apoproteins of Bacillus polymyxa ferredoxin and adrenodoxin. Detection and quantitation are accomplished by electron paramagnetic resonance of reduced proteins at low temperatures. Both procedures clearly show that succinate dehydrogenase contains two dimeric (Fe2S2) and one tetrameric (Fe4S4) centers per mole of histidyl flavin, accounting for all eight nonheme iron and eight labile sulfur atoms found by chemical analysis. These results remove uncertainties created by the less than stoichiometric amounts of binuclear centers detected by electron paramagnetic resonance after dithionite reduction and provide secure characterization of the iron-sulfur centers in this enzyme.
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页码:3805 / 3808
页数:4
相关论文
共 29 条
[1]   EPR SIGNAL INTENSITY AND POWDER SHAPES - RE-EXAMINATION [J].
AASA, R ;
VANNGARD, T .
JOURNAL OF MAGNETIC RESONANCE, 1975, 19 (03) :308-315
[2]  
ACKRELL BAC, 1977, J BIOL CHEM, V252, P6963
[3]   DISPLACEMENT OF IRON-SULFUR CLUSTERS FROM FERREDOXINS AND OTHER IRON-SULFUR PROTEINS [J].
AVERILL, BA ;
BALE, JR ;
ORMEJOHNSON, WH .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1978, 100 (10) :3034-3043
[4]  
BALE JR, 1974, THESIS U WISCONSIN
[5]   STUDIES ON SUCCINATE-DEHYDROGENASE .30. INTERRELATIONS OF RECONSTITUTION ACTIVITY, REACTIONS WITH ELECTRON-ACCEPTORS, AND IRON-SULFUR CENTERS IN SUCCINATE-DEHYDROGENASE [J].
BEINERT, H ;
ACKRELL, BAC ;
VINOGRADOV, AD ;
KEARNEY, EB ;
SINGER, TP .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1977, 182 (01) :95-106
[6]   IRON-SULFUR COMPONENTS OF SUCCINATE-DEHYDROGENASE - STOICHIOMETRY AND KINETIC-BEHAVIOR IN ACTIVATED PREPARATIONS [J].
BEINERT, H ;
ACKRELL, BAC ;
KEARNEY, EB ;
SINGER, TP .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1975, 54 (01) :185-194
[7]  
Brumby P. E., 1967, METHOD ENZYMOL, V10, P463, DOI DOI 10.1016/0076-6879(67)10078-5
[8]  
COLES CJ, 1972, PHYSIOL CHEM PHYS M, V4, P301
[9]  
DAVIS KA, 1971, BIOCHEMISTRY-US, V10, P2509
[10]   CHEMICAL CHARACTERIZATION OF HIGH POTENTIAL IRON PROTEINS FROM CHROMATIUM AND RHODOPSEUDOMONAS GELATINOSA [J].
DUS, K ;
DEKLERK, H ;
BARTSCH, RG ;
SLETTEN, K .
BIOCHIMICA ET BIOPHYSICA ACTA, 1967, 140 (02) :291-&