PDBsum entry 1anh

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Hydrolase PDB id
Protein chains
449 a.a.
PO4 ×2
_ZN ×6
Waters ×82
Superseded by: 2anh 2anh
PDB id:
Name: Hydrolase
Structure: Alkaline phosphatase mutation with asp 153 replaced by his (d153h) complexed with zinc and po4
Source: (Escherichia coli) strain: ek1457, expression system: (escherichia coli) sm547, plasmid: pek175, gene: phoa
Authors: J.E.Murphy,X.Xu,E.R.Kantrowitz
Key ref: J.E.Murphy et al. (1993). Conversion of a magnesium binding site into a zinc binding site by a single amino acid substitution in Escherichia coli alkaline phosphatase. J Biol Chem, 268, 21497-21500. PubMed id: 8407998
19-Oct-94     Release date:   26-Jan-95    
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Protein chains
No UniProt id for this chain
Struc: 449 a.a.
Key:    Secondary structure  CATH domain


J Biol Chem 268:21497-21500 (1993)
PubMed id: 8407998  
Conversion of a magnesium binding site into a zinc binding site by a single amino acid substitution in Escherichia coli alkaline phosphatase.
J.E.Murphy, X.Xu, E.R.Kantrowitz.
The replacement of aspartic acid by histidine at position 153 in Escherichia coli alkaline phosphatase results in a mutant enzyme that is remarkably similar to certain mammalian alkaline phosphatases that are activated by magnesium in a time-dependent fashion. These mammalian alkaline phosphatases have histidine at the position corresponding to 153 of the E. coli sequence. Here we report the three-dimensional structure of the mutant E. coli alkaline phosphatase with histidine at position 153. The structure reveals that the octahedral magnesium binding site has been converted to a tetrahedral zinc binding site with an imidazole ring nitrogen of His-153 as one of the ligands to the zinc. The alteration in metal binding caused by the mutation could explain the origin of the magnesium activation observed with the mammalian alkaline phosphatases. The structure also reveals differences in the mode of phosphate binding, explaining the enhanced phosphate affinity and the reduced activity of the mutant enzyme in the presence of zinc.

Literature references that cite this PDB file's key reference

  PubMed id Reference
  19916164 D.Koutsioulis, A.Lyskowski, S.Mäki, E.Guthrie, G.Feller, V.Bouriotis, and P.Heikinheimo (2010).
Coordination sphere of the third metal site is essential to the activity and metal selectivity of alkaline phosphatases.
  Protein Sci, 19, 75-84.
PDB codes: 2w5v 2w5w 2w5x
19272175 S.L.Wu, C.C.Li, J.C.Chen, Y.J.Chen, C.T.Lin, T.Y.Ho, and C.Y.Hsiang (2009).
Mutagenesis identifies the critical amino acid residues of human endonuclease G involved in catalysis, magnesium coordination, and substrate specificity.
  J Biomed Sci, 16, 6.  
17989875 J.Li, L.Xu, and F.Yang (2007).
Expression and characterization of recombinant thermostable alkaline phosphatase from a novel thermophilic bacterium Thermus thermophilus XM.
  Acta Biochim Biophys Sin (Shanghai), 39, 844-850.  
15885097 T.Harada, I.Koyama, T.Matsunaga, A.Kikuno, T.Kasahara, M.Hassimoto, D.H.Alpers, and T.Komoda (2005).
Characterization of structural and catalytic differences in rat intestinal alkaline phosphatase isozymes.
  FEBS J, 272, 2477-2486.  
15725663 Y.Suzuki, Y.Mizutani, T.Tsuji, N.Ohtani, K.Takano, M.Haruki, M.Morikawa, and S.Kanaya (2005).
Gene cloning, overproduction, and characterization of thermolabile alkaline phosphatase from a psychrotrophic bacterium.
  Biosci Biotechnol Biochem, 69, 364-373.  
15333925 Backer, S.McSweeney, P.F.Lindley, and E.Hough (2004).
Ligand-binding and metal-exchange crystallographic studies on shrimp alkaline phosphatase.
  Acta Crystallogr D Biol Crystallogr, 60, 1555-1561.
PDB codes: 1shn 1shq
11910033 C.L.Wojciechowski, J.P.Cardia, and E.R.Kantrowitz (2002).
Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity.
  Protein Sci, 11, 903-911.  
11342026 R.Q.Zhang, Q.X.Chen, R.Xiao, L.P.Xie, X.G.Zeng, and H.M.Zhou (2001).
Inhibition kinetics of green crab (Scylla serrata) alkaline phosphatase by zinc ions: a new type of complexing inhibition.
  Biochim Biophys Acta, 1545, 6.  
11571149 S.Zappa, J.L.Rolland, D.Flament, Y.Gueguen, J.Boudrant, and J.Dietrich (2001).
Characterization of a highly thermostable alkaline phosphatase from the euryarchaeon Pyrococcus abyssi.
  Appl Environ Microbiol, 67, 4504-4511.  
10672035 M.Rina, C.Pozidis, K.Mavromatis, M.Tzanodaskalaki, M.Kokkinidis, and V.Bouriotis (2000).
Alkaline phosphatase from the Antarctic strain TAB5. Properties and psychrophilic adaptations.
  Eur J Biochem, 267, 1230-1238.  
10940645 Q.X.Chen, W.Z.Zheng, J.Y.Lin, Y.Shi, W.Z.Xie, and H.M.Zhou (2000).
Effect of metal ions on the activity of green crab (Scylla serrata) alkaline phosphatase.
  Int J Biochem Cell Biol, 32, 879-885.  
10924156 S.H.Francis, I.V.Turko, K.A.Grimes, and J.D.Corbin (2000).
Histidine-607 and histidine-643 provide important interactions for metal support of catalysis in phosphodiesterase-5.
  Biochemistry, 39, 9591-9596.  
10584076 M.Bortolato, F.Besson, and B.Roux (1999).
Role of metal ions on the secondary and quaternary structure of alkaline phosphatase from bovine intestinal mucosa.
  Proteins, 37, 310-318.  
10556703 T.Park, J.H.Lee, H.K.Kim, H.S.Hoe, and S.T.Kwon (1999).
Nucleotide sequence of the gene for alkaline phosphatase of Thermus caldophilus GK24 and characteristics of the deduced primary structure of the enzyme.
  FEMS Microbiol Lett, 180, 133-139.  
7541135 C.A.Brennan, K.Christianson, M.A.La Fleur, and W.Mandecki (1995).
A molecular sensor system based on genetically engineered alkaline phosphatase.
  Proc Natl Acad Sci U S A, 92, 5783-5787.  
  8520475 L.Ma, T.T.Tibbitts, and E.R.Kantrowitz (1995).
Escherichia coli alkaline phosphatase: X-ray structural studies of a mutant enzyme (His-412-->Asn) at one of the catalytically important zinc binding sites.
  Protein Sci, 4, 1498-1506.
PDB codes: 1ali 1alj
  7703848 T.T.Tibbitts, X.Xu, and E.R.Kantrowitz (1994).
Kinetics and crystal structure of a mutant Escherichia coli alkaline phosphatase (Asp-369-->Asn): a mechanism involving one zinc per active site.
  Protein Sci, 3, 2005-2014.
PDB code: 1alh
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