Structures of Selenomonas ruminantium phytase in complex with persulfated phytate:: DSP phytase fold and mechanism for sequential substrate hydrolysis

被引:78
作者
Chu, HM
Guo, RT
Lin, TW
Chou, CC
Shr, HL
Lai, HL
Tang, TY
Cheng, KJ
Selinger, BL
Wang, AHJ [1 ]
机构
[1] Natl Taiwan Univ, Inst Biochem Sci, Taipei 106, Taiwan
[2] Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan
[3] Acad Sinica, Core Facil Protein X Ray Crystallog, Taipei 115, Taiwan
[4] Acad Sinica, Inst BioAgr Sci, Taipei 115, Taiwan
[5] Univ Lethbridge, Dept Biol Sci, Lethbridge, AB T1K 3M4, Canada
[6] Acad Sinica, Taiwan Int Grad Program, Taipei 115, Taiwan
关键词
D O I
10.1016/j.str.2004.08.010
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
Various inositide phosphatases participate in the regulation of inositol polyphosphate signaling molecules. Plant phytases are phosphatases that hydrolyze phytate to less-phosphorylated myo-inositol derivatives and phosphate. The phytase from Selenomonas ruminantium shares no sequence homology with other microbial phytases. Its crystal structure revealed a phytase fold of the dual-specificity phosphatase type. The active site is located near a conserved cysteine-containing (Cys241) P loop. We also solved two other crystal forms in which an inhibitor, myo-inositol hexasulfate, is cocrystallized with the enzyme. In the "standby" and the "inhibited" crystal forms, the inhibitor is bound, respectively, in a pocket slightly away from Cys241 and at the substrate binding site where the phosphate group to be hydrolyzed is held close to the-SH group of Cys2411. Our structural and mutagenesis studies allow us to visualize the way in which the P loop-containing phytase attracts and hydrolyzes the substrate (phytate) sequentially.
引用
收藏
页码:2015 / 2024
页数:10
相关论文
共 37 条
[1]   STRUCTURE OF INOSITOL HEXAPHOSPHATE-HUMAN DEOXYHEMOGLOBIN COMPLEX [J].
ARNONE, A ;
PERUTZ, MF .
NATURE, 1974, 249 (5452) :34-36
[2]   A novel staining method for detecting phytase activity [J].
Bae, HD ;
Yanke, LJ ;
Cheng, KJ ;
Selinger, LB .
JOURNAL OF MICROBIOLOGICAL METHODS, 1999, 39 (01) :17-22
[3]  
Brunger AT, 1998, ACTA CRYSTALLOGR D, V54, P905, DOI 10.1107/s0907444998003254
[4]   The human and rat forms of multiple inositol polyphosphate phosphatase: functional homology with a histidine acid phosphatase up-regulated during endochondral ossification [J].
Caffrey, JJ ;
Hidaka, K ;
Matsuda, M ;
Hirata, M ;
Shears, SB .
FEBS LETTERS, 1999, 442 (01) :99-104
[5]   Structure and mechanism of the RNA triphosphatase component of mammalian mRNA capping enzyme [J].
Changela, A ;
Ho, CK ;
Martins, A ;
Shuman, S ;
Mondragón, A .
EMBO JOURNAL, 2001, 20 (10) :2575-2586
[6]   A CATALYTIC MECHANISM FOR THE DUAL-SPECIFIC PHOSPHATASES [J].
DENU, JM ;
DIXON, JE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (13) :5910-5914
[7]   Visualization of intermediate and transition-state structures in protein-tyrosine phosphatase catalysis [J].
Denu, JM ;
Lohse, DL ;
Vijayalakshmi, J ;
Saper, MA ;
Dixon, JE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (06) :2493-2498
[8]   Protein tyrosine phosphatases: mechanisms of catalysis and regulation [J].
Denu, JM ;
Dixon, JE .
CURRENT OPINION IN CHEMICAL BIOLOGY, 1998, 2 (05) :633-641
[9]   Production of selenomethionine-labelled proteins using simplified culture conditions and generally applicable host/vector systems [J].
Guerrero, SA ;
Hecht, HJ ;
Hofmann, B ;
Biebl, H ;
Singh, M .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2001, 56 (5-6) :718-723
[10]   SWISS-MODEL and the Swiss-PdbViewer: An environment for comparative protein modeling [J].
Guex, N ;
Peitsch, MC .
ELECTROPHORESIS, 1997, 18 (15) :2714-2723