NONSELENIUM GLUTATHIONE-PEROXIDASE WITHOUT GLUTATHIONE S-TRANSFERASE ACTIVITY FROM BOVINE CILIARY BODY

被引:59
作者
SHICHI, H [1 ]
DEMAR, JC [1 ]
机构
[1] OAKLAND UNIV,EYE RES INST,ROCHESTER,MI 48063
关键词
bovine; ciliary body; glutathione peroxidase; properties; purification; selenium-independent;
D O I
10.1016/0014-4835(90)90040-2
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
A glutathione peroxidase was purified from bovine ciliary body by ammonium sulfate fractionation. Sephacryl S-300 gel filtration, diethylaminoethyl (DEAE)-cellulose chromatography and hydroxyapatite chromatography. The purified enzyme has an apparent mw of 112 kDa by gel filtration and 29 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme therefore is composed of four identical subunits. The ciliary enzyme is active with H2O2 (25), cumene hydroperoxide (170), t-butyl hydroperoxide (22), triphenylcarbinyl hydroperoxide (12), linoleic hydroproxide (34) and 5-phenylpentenyl hydroperoxide (22): the numbers after substrates are K′m in μm. Glutathione is essential for the reaction; l-cysteine, dithiothreitol and 2-mercaptoethanol are inactive. Mercaptosuccinate (10 μm) inhibits the enzyme competitively (Ki = 7 μM) when cumene hydroperoxide is substrate, and uncompetitively (Ki = 10 μM) when H2O2 is substrate. No selenium was found in the enzyme by the fluorometric assay with 2,3-diaminonaphthalene. The enzyme demonstrates no glutathione S-transferase activity when tested with 1-chloro-2,4-dinitrobenzene, and several other compounds. A partial sequence of the enzyme shows some similarities both to Se-glutathione peroxidases and a glutathione S-transferase isozyme. © 1990.
引用
收藏
页码:513 / 520
页数:8
相关论文
共 27 条
[1]   THE DISTRIBUTION OF PEROXIDE REGULATING ENZYMES IN THE CANINE EYE [J].
ARMSTRONG, D ;
SANTANGELO, G ;
CONNOLE, E .
CURRENT EYE RESEARCH, 1981, 1 (04) :225-242
[2]   IMMUNOHISTOCHEMICAL LOCALIZATION OF GLUTATHIONE-PEROXIDASE IN OCULAR TISSUE [J].
ATALLA, LR ;
SEVANIAN, A ;
RAO, NA .
CURRENT EYE RESEARCH, 1988, 7 (10) :1023-1027
[3]   PH CONTROL IN THE FLUOROMETRIC ASSAY FOR SELENIUM WITH 2,3-DIAMINONAPHTHALENE [J].
BAYFIELD, RF ;
ROMALIS, LF .
ANALYTICAL BIOCHEMISTRY, 1985, 144 (02) :569-576
[4]   GLUTATHIONE-PEROXIDASE FROM BOVINE LENS - A SELENOENZYME [J].
BERGAD, PL ;
RATHBUN, WB ;
LINDER, W .
EXPERIMENTAL EYE RESEARCH, 1982, 34 (01) :131-144
[5]   MOLECULAR MECHANISM OF CATARACTOGENESIS .3. TOXIC METABOLITES OF OXYGEN AS INITIATORS OF LIPID-PEROXIDATION AND CATARACT [J].
BHUYAN, KC ;
BHUYAN, DK .
CURRENT EYE RESEARCH, 1984, 3 (01) :67-81
[6]  
CHAUDIERE J, 1984, J BIOL CHEM, V259, P1043
[7]  
DAS ND, 1981, EXP EYE RES, V29, P109
[8]   GLUTATHIONE PEROXIDASE .1. PURIFICATION AND ESTIMATION OF MOLECULAR WEIGHT [J].
FLOHE, L ;
EISELE, B ;
WENDEL, A .
HOPPE-SEYLERS ZEITSCHRIFT FUR PHYSIOLOGISCHE CHEMIE, 1971, 352 (02) :151-&
[9]   THE AMINO-ACID-SEQUENCE OF BOVINE GLUTATHIONE-PEROXIDASE [J].
GUNZLER, WA ;
STEFFENS, GJ ;
GROSSMANN, A ;
KIM, SMA ;
OTTING, F ;
WENDEL, A ;
FLOHE, L .
HOPPE-SEYLERS ZEITSCHRIFT FUR PHYSIOLOGISCHE CHEMIE, 1984, 365 (02) :195-212
[10]  
HABIG WH, 1974, J BIOL CHEM, V249, P7130