ElaC encodes a novel binuclear zinc phosphodiesterase

被引:75
作者
Vogel, A
Schilling, O
Niecke, M
Bettmer, J
Meyer-Klaucke, W
机构
[1] European Mol Biol Lab Outstn Hamburg, D-22603 Hamburg, Germany
[2] Univ Hamburg, Dept Expt Phys, D-22761 Hamburg, Germany
[3] Johannes Gutenberg Univ Mainz, Dept Analyt Chem, D-55099 Mainz, Germany
关键词
D O I
10.1074/jbc.M112047200
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
ElaC is a widespread gene found in eubacteria, archaebacteria, and mammals with a highly conserved sequence. Two human ElaC variants were recently associated with cancer (Tavtigian, S. V., Simard, J., Teng, D. H., Abtin, V., Baumgard, M., Beck, A., Camp, N. J., Carillo, A. R., Chen, Y., Dayananth, P., Desrochers, M., Dumont, M., Farnham, J. M., Frank, D., Frye, C., Ghaffari, S., Gupte, J. S., Hu, R., Iliev, D., Janecki, T., Kort, E. N., Laity, F, E., Leavitt, A., Leblanc, G., McArthur-Morrison, J., Pederson, A., Penn, B., Peterson, K. T., Reid, J. E., Richards, S., Schroeder, M., Smith, R., Snyder, S. C., Swedlund, B., Swensen, J., Thomas, A., Tranchant, M., Woodland, A. M., Labrie, F., Skolnick, M. H., Neuhausen, S., Rommens, J., and Cannon-Albright, L. A. (2001) Nat. Genet. 27, 172-180; Yanaihara, N., Kohno, T., Takakura, S., Takei, M, Otsuka, A., Sunaga, N., Takahashi, M., Yamazaki, M., Tashiro, H., Fukuzumi, Y., Fujimori, Y., Hagiwara, K., Tanaka, T., and Yokota, J. (2001) Genomics 72, 169-179). Analysis of the primary sequence indicates homology to an arylsulfatase and predicts a metallo-beta-lactamase fold. At present, no ElaC gene product has been investigated. We cloned the Escherichia coli ElaC gene and purified the recombinant gene product. An enzymatic analysis showed that ElaC does not encode an arylsulfatase but rather encodes a phosphodiesterase that hydrolyzes bis(p-nitrophenyl)phosphate with a k(cat) of 59 s(-1) and K' of 4 mM. Kinetic analysis of the dimeric enzyme revealed positive cooperativity for the substrate bis(p-nitrophenyl)phosphate with a Hill coefficient of 1.6, whereas hydrolysis of the substrate thymidine-5'-p-nitrophenyl phosphate followed Michaelis-Menten kinetics. Furthermore, the enzyme is capable of binding two zinc or two iron ions. However, it displays phosphodiesterase activity only in the zinc form. The metal environment characterized by zinc K-edge x-ray absorption spectroscopy was modeled with two histidine residues, one carboxylate group, and 1.5 oxygen atoms. This corresponds to the coordination found in other metallo-beta-lactamase domain proteins. Phosphodiesterase activity is strongly dependent on the presence of zinc. These results identify the currently unassigned gene product ElaC to be a novel binuclear zinc phosphodiesterase.
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页码:29078 / 29085
页数:8
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共 38 条
[1]   Analysis of zinc binding sites in protein crystal structures [J].
Alberts, IL ;
Nadassy, K ;
Wodak, SJ .
PROTEIN SCIENCE, 1998, 7 (08) :1700-1716
[2]  
Aravind L, 1999, In Silico Biol, V1, P69
[3]   ARYLSULFATASE FROM ALTEROMONAS-CARRAGEENOVORA [J].
BARBEYRON, T ;
POTIN, P ;
RICHARD, C ;
COLLIN, O ;
KLOAREG, B .
MICROBIOLOGY-UK, 1995, 141 :2897-2904
[4]   CONSTRAINED AND RESTRAINED REFINEMENT IN EXAFS DATA-ANALYSIS WITH CURVED WAVE THEORY [J].
BINSTED, N ;
STRANGE, RW ;
HASNAIN, SS .
BIOCHEMISTRY, 1992, 31 (48) :12117-12125
[5]   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
[6]   THE 3-D STRUCTURE OF A ZINC METALLO-BETA-LACTAMASE FROM BACILLUS-CEREUS REVEALS A NEW-TYPE OF PROTEIN FOLD [J].
CARFI, A ;
PARES, S ;
DUEE, E ;
GALLENI, M ;
DUEZ, C ;
FRERE, JM ;
DIDEBERG, O .
EMBO JOURNAL, 1995, 14 (20) :4914-4921
[7]   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
[8]   Expansion of the zinc metallo-hydrolase family of the β-lactamase fold [J].
Daiyasu, H ;
Osaka, K ;
Ishino, Y ;
Toh, H .
FEBS LETTERS, 2001, 503 (01) :1-6
[9]   Identification, characterization, and cloning of a phosphonate monoester hydrolase from Burkholderia caryophilli PG2982 [J].
Dotson, SB ;
Smith, CE ;
Ling, CS ;
Barry, GF ;
Kishore, GM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (42) :25754-25761
[10]  
Frazao C, 2000, NAT STRUCT BIOL, V7, P1041