Catalytic and structural function of zinc for the hydantoinase from Arthrobacter aurescens DSM 3745

被引:26
作者
May, O
Siemann, M
Siemann, MG
Syldatk, C
机构
[1] Univ Stuttgart, Inst Bioverfahrenstech, D-70569 Stuttgart, Germany
[2] Tech Univ Clausthal, Inst Mineral, Abt Geochem, D-38678 Clausthal Zellerfeld, Germany
关键词
hydantoinase; cyclic amidase; Arthrobacter aurescens; metalloenzyme; zinc;
D O I
10.1016/S1381-1177(97)00038-6
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
Metal dependency of the hydantoin amidohydrolase (hydantoinase) from Arthrobacter aurescens DSM 3745 has been analyzed based on kinetic studies of metal/chelator-caused enzyme inactivation, denaturation and reactivation, accompanied by the identification of specific metal binding ligands. The enzyme can be inactivated by metal chelating agents and-apart from the loss of its activity-completely dissociates into its subunits. Enzyme activity can be restored from recollected monomers by the addition of cobalt, manganese or zinc-ions, whereas nickel and magnesia remain ineffective. Subjection of the hydantoinase to metal analysis reveals a content of 10 mol zinc per mol enzyme. Zinc plays an essential role not only for the catalytic activity but also for the stabilization of the active quarternary structure of the hydantoinase. Histidine-specific chemical modification of the enzyme causes a complete loss of the catalytic activity and reveals histidine residues as putative zinc binding ligands. Both, the metal/chelator-caused enzyme inactivation as well as the metal-caused enzyme reactivation, can be reduced in the presence of the substrate. Therefore, it is very likely that at least one metal-ion acts specifically near or at the active site of the enzyme. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:211 / 218
页数:8
相关论文
共 22 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   BOVINE LIVER DIHYDROPYRIMIDINE AMIDOHYDROLASE - PURIFICATION, PROPERTIES, AND CHARACTERIZATION AS A ZINC METALLOENZYME [J].
BROOKS, KP ;
JONES, EA ;
KIM, BD ;
SANDER, EG .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1983, 226 (02) :469-483
[3]  
CHRISTIANSON DW, 1991, ADV PROTEIN CHEM, V42, P281
[4]  
Holm L, 1997, PROTEINS, V28, P72, DOI 10.1002/(SICI)1097-0134(199705)28:1<72::AID-PROT7>3.0.CO
[5]  
2-L
[6]   ACID-BASE CATALYTIC MECHANISM OF DIHYDROPYRIMIDINASE FROM PH STUDIES [J].
JAHNKE, K ;
PODSCHUN, B ;
SCHNACKERZ, KD ;
KAUTZ, J ;
COOK, PF .
BIOCHEMISTRY, 1993, 32 (19) :5160-5166
[7]   PURIFICATION AND PROPERTIES OF 5,6-DIHYDROPYRIMIDINE AMIDOHYDROLASE FROM CALF LIVER [J].
KAUTZ, J ;
SCHNACKERZ, KD .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1989, 181 (02) :431-435
[8]   Crystallization and preliminary X-ray analysis of a hydantoinase from Arthrobacter aurescens DSM 3745 [J].
May, O ;
Siemann, M ;
Syldatk, C ;
Niefind, K ;
Schomburg, D .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1996, 52 :1209-1210
[9]  
MAY O, 1997, UNPUB J MOL CATAL B
[10]  
MAY O, 1997, UNPUB J BACTERIOL