IDENTIFICATION OF LYS116 AS THE TARGET OF N-ETHYLMALEIMIDE INACTIVATION OF FERREDOXIN - NADP+ OXIDOREDUCTASE

被引:17
作者
ALIVERTI, A
GADDA, G
RONCHI, S
ZANETTI, G
机构
[1] UNIV MILAN, DIPARTIMENTO FISIOL & BIOCHIM GEN, VIA CELORIA 26, I-20133 MILAN, ITALY
[2] UNIV MILAN, IST FISIOL VET & BIOCHIM, I-20133 MILAN, ITALY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1991年 / 198卷 / 01期
关键词
D O I
10.1111/j.1432-1033.1991.tb15981.x
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
Oxidized ferredoxin: NADP+ oxidoreductase (FNR) was slowly and irreversibly inactivated by N-ethylmaleimide. Complete protection against inactivation was afforded by saturating concentrations of NADP+. In the presence of NADPH, a rapid inhibition of the enzyme ensued; however, this inhibition was found to be reversible. In the tryptic map of the flavoprotein, modified with N-ethyl[2,3-C-14]maleimide in oxidizing conditions, a unique radioactive peptide was found. Its sequence comprised residues 110-117 of the enzyme; Lys116 was shown to be the residue alkylated by N-ethylmaleimide. It is noteworthy that the same residue of FNR was found to be modified by 5-dimethylaminoaphthalene-1-sulfonyl(dansyl) chloride at the putative NADP(H)-binding site [Cidaria, D., Biondi, P. A., Zanetti, G. & Ronchi, S. (1985) Eur. J. Biochem. 146, 295-299]. Furthermore, the data reported here demonstrate that the sulfhydryl groups of FNR are not involved in enzyme inactivation by N-ethylmaleimide.
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页码:21 / 24
页数:4
相关论文
共 18 条
[1]   A TPNH DIAPHORASE FROM CHLOROPLASTS [J].
ABRAMSKY, MA ;
JAGENDORF, AT .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1956, 65 (02) :475-490
[2]   EXPRESSION IN ESCHERICHIA-COLI OF FERREDOXIN - NADP+ REDUCTASE FROM SPINACH - BACTERIAL SYNTHESIS OF THE HOLOFLAVOPROTEIN AND OF AN ACTIVE ENZYME FORM LACKING THE 1ST 28 AMINO-ACID-RESIDUES OF THE SEQUENCE [J].
ALIVERTI, A ;
JANSEN, T ;
ZANETTI, G ;
RONCHI, S ;
HERRMANN, RG ;
CURTI, B .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 191 (03) :551-555
[3]   EVIDENCE FOR POSSIBLE NONSPECIFIC REACTIONS BETWEEN N-ETHYLMALEIMIDE AND PROTEINS [J].
BREWER, CF ;
RIEHM, JP .
ANALYTICAL BIOCHEMISTRY, 1967, 18 (02) :248-&
[4]   ISOLATION AND SEQUENCING OF AN ACTIVE-SITE PEPTIDE FROM SPINACH FERREDOXIN-NADP+ OXIDOREDUCTASE AFTER AFFINITY LABELING WITH PERIODATE-OXIDIZED NADP+ [J].
CHAN, RL ;
CARRILLO, N ;
VALLEJOS, RH .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1985, 240 (01) :172-177
[5]   THE NADP+-BINDING SITE OF FERRODOXIN-NADP+ REDUCTASE - SEQUENCE OF THE PEPTIDE CONTAINING THE ESSENTIAL LYSINE RESIDUE [J].
CIDARIA, D ;
BIONDI, PA ;
ZANETTI, G ;
RONCHI, S .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1985, 146 (02) :295-299
[6]   EVIDENCE FOR ROLE OF SULFHYDRYL-GROUPS IN A PH-DEPENDENT TRANSITION OF FERREDOXIN=NADP OXIDOREDUCTASE [J].
DAVIS, DJ ;
SANPIETRO, A .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1977, 184 (02) :572-577
[7]  
FORTI G, 1967, BIOCH CHLOROPLASTS, V2, P523
[8]  
GADDA G, 1990, J BIOL CHEM, V265, P11955
[9]   A PEPTIDE CONTAINING A REACTIVE LYSYL GROUP FROM OX LIVER GLUTAMATE DEHYDROGENASE [J].
HOLBROOK, JJ ;
JECKEL, R .
BIOCHEMICAL JOURNAL, 1969, 111 (05) :689-+
[10]   AMINO-ACID SEQUENCE OF SPINACH FERREDOXIN - NADP+ OXIDOREDUCTASE [J].
KARPLUS, PA ;
WALSH, KA ;
HERRIOTT, JR .
BIOCHEMISTRY, 1984, 23 (26) :6576-6583