Arabidopsis glyoxalase II contains a zinc/iron binuclear metal center that is essential for substrate binding and catalysis

被引:74
作者
Zang, TM [1 ]
Hollman, DA [1 ]
Crawford, PA [1 ]
Crowder, MW [1 ]
Makaroff, CA [1 ]
机构
[1] Miami Univ, Dept Chem & Biochem, Oxford, OH 45056 USA
关键词
D O I
10.1074/jbc.M005090200
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
Glyoxalase II participates in the cellular detoxification of cytotoxic and mutagenic 2-oxoaldehydes. Because of its role in chemical detoxification, glyoxalase II has been studied as a potential anti-cancer and/or antiprotozoal target; however, very little is known about the active site and reaction mechanism of this important enzyme. To characterize the active site and kinetic mechanism of the enzyme, a detailed mutational study of Arabidopsis glyoxalase II was conducted. Data presented here demonstrate for the first time that the cytoplasmic form of Arabidopsis glyoxalase II contains an iron-zinc binuclear metal center that is essential for activity. Both metals participate in substrate binding, transition state stabilization, and the hydrolysis reaction. Subtle alterations in the geometry and/or electrostatics of the binuclear center have profound effects on the activity of the enzyme. Additional residues important in substrate binding have also been identified. An overall reaction mechanism for glyoxalase II is proposed based on the mutational and kinetic data from this study and crystallographic data on human glyoxalase II. Information presented here provides new insights into the active site and reaction mechanism of glyoxalase II that can be used for the rational design of glyoxalase II inhibitors.
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页码:4788 / 4795
页数:8
相关论文
共 41 条
[1]   INHIBITORS OF GLYOXALASE-I - DESIGN, SYNTHESIS, INHIBITORY CHARACTERISTICS AND BIOLOGICAL EVALUATION [J].
ALLEN, RE ;
LO, TWC ;
THORNALLEY, PJ .
BIOCHEMICAL SOCIETY TRANSACTIONS, 1993, 21 (02) :535-540
[2]   PURIFICATION AND CHARACTERIZATION OF GLYOXALASE-II FROM HUMAN RED-BLOOD-CELLS [J].
ALLEN, RE ;
THEODORE, WCL ;
THORNALLEY, PJ .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 213 (03) :1261-1267
[3]   S-2-HYDROXYACYLGLUTATHIONE HYDROLASE (GLYOXALASE-II) - ACTIVE-SITE MAPPING OF A NONSERINE THIOLESTERASE [J].
BALL, JC ;
VANDERJAGT, DL .
BIOCHEMISTRY, 1981, 20 (04) :899-905
[4]   Crystal structure of human glyoxalase II and its complex with a glutathione thiolester substrate analogue [J].
Cameron, AD ;
Ridderström, M ;
Olin, B ;
Mannervik, B .
STRUCTURE, 1999, 7 (09) :1067-1078
[5]   Catalysis by metal-activated hydroxide in zinc and manganese metalloenzymes [J].
Christianson, DW ;
Cox, JD .
ANNUAL REVIEW OF BIOCHEMISTRY, 1999, 68 :33-57
[6]   Crystal structure of the wide-spectrum binuclear zinc beta-lactamase from Bacteroides fragilis [J].
Concha, NO ;
Rasmussen, BA ;
Bush, K ;
Herzberg, O .
STRUCTURE, 1996, 4 (07) :823-836
[7]   OPTIMIZATION OF EFFICIENCY IN THE GLYOXALASE PATHWAY [J].
CREIGHTON, DJ ;
MIGLIORINI, M ;
POURMOTABBED, T ;
GUHA, MK .
BIOCHEMISTRY, 1988, 27 (19) :7376-7384
[8]   Glyoxalase II from A-thaliana requires Zn(II) for catalytic activity [J].
Crowder, MW ;
Maiti, MK ;
Banovic, L ;
Makaroff, CA .
FEBS LETTERS, 1997, 418 (03) :351-354
[9]   Characterization of the metal-binding sites of the beta-lactamase from Bacteroides fragilis [J].
Crowder, MW ;
Wang, ZG ;
Franklin, SL ;
Zovinka, EP ;
Benkovic, SJ .
BIOCHEMISTRY, 1996, 35 (37) :12126-12132
[10]  
CROWDER MW, 1999, ANTIMICROBIAL AGEN 1, V3, P105