PDBsum entry 1ajd

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protein metals Protein-protein interface(s) links
Non specific mono-esterase PDB id
Protein chains
449 a.a. *
_MG ×2
_ZN ×4
Waters ×4
* Residue conservation analysis
PDB id:
Name: Non specific mono-esterase
Title: Three-dimensional structure of the d153g mutant of e. Coli alkaline phosphatase: a mutant with weaker magnesium binding and increased catalytic activity
Structure: Alkaline phosphatase intermediate ii of holo enzyme. Chain: a, b. Engineered: yes. Mutation: yes. Other_details: mutant with 4-fold increased activity and weaker mg binding, structure solved after resoaking metal ions for two months into the apo crystals
Source: Escherichia coli. Organism_taxid: 562. Gene: phoa. Expressed in: escherichia coli. Expression_system_taxid: 562. Restriction fragment. Other_details: lac promoter
Biol. unit: Dimer (from PQS)
2.50Å     R-factor:   0.162    
Authors: C.G.Dealwis,L.Chen,C.Abad-Zapatero
Key ref:
C.G.Dealwis et al. (1995). Crystallographic analysis of reversible metal binding observed in a mutant (Asp153-->Gly) of Escherichia coli alkaline phosphatase. Biochemistry, 34, 13967-13973. PubMed id: 7577993 DOI: 10.1021/bi00043a001
18-Aug-95     Release date:   14-Nov-95    
Go to PROCHECK summary

Protein chains
Pfam   ArchSchema ?
P00634  (PPB_ECOLI) -  Alkaline phosphatase
471 a.a.
449 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.  - Alkaline phosphatase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: A phosphate monoester + H2O = an alcohol + phosphate
phosphate monoester
+ H(2)O
= alcohol
+ phosphate
      Cofactor: Magnesium; Zinc
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     periplasmic space   2 terms 
  Biological process     metabolic process   3 terms 
  Biochemical function     catalytic activity     10 terms  


DOI no: 10.1021/bi00043a001 Biochemistry 34:13967-13973 (1995)
PubMed id: 7577993  
Crystallographic analysis of reversible metal binding observed in a mutant (Asp153-->Gly) of Escherichia coli alkaline phosphatase.
C.G.Dealwis, C.Brennan, K.Christianson, W.Mandecki, C.Abad-Zapatero.
Here we present the refined crystal structures of three different conformational states of the Asp153-->Gly mutant (D153G) of alkaline phosphatase (AP), a metalloenzyme from Escherichia coli. The apo state is induced in the crystal over a 3 month period by metal depletion of the holoenzyme crystals. Subsequently, the metals are reintroduced in the crystalline state in a time-dependent reversible manner without physically damaging the crystals. Two structural intermediates of the holo form based on data from a 2 week (intermediate I) and a 2 month soak (intermediate II) of the apo crystals with Mg2+ and Zn2+ have been identified. The three-dimensional crystal structures of the apo (R = 18.1%), intermediate I (R = 19.5%), and intermediate II (R = 19.9%) of the D153G enzyme have been refined and the corresponding structures analyzed and compared. Large conformational changes that extend from the mutant active site to surface loops, located 20 A away, are observed in the apo structure with respect to the holo structure. The structure of intermediate I shows the recovery of the entire enzyme to an almost native-like conformation, with the exception of residues Asp 51 and Asp 369 in the active site and the surface loop (406-410) which remains partially disordered. In the three-dimensional structure of intermediate II, both Asp 51 and Asp 369 are essentially in a native-like conformation, but the main chain of residues 406-408 within the loop is still not fully ordered. The D153G mutant protein exhibits weak, reversible, time dependent metal binding in solution and in the crystalline state.(ABSTRACT TRUNCATED AT 250 WORDS)

Literature references that cite this PDB file's key reference

  PubMed id Reference
16815919 P.Llinas, M.Masella, T.Stigbrand, A.Ménez, E.A.Stura, and M.H.Le Du (2006).
Structural studies of human alkaline phosphatase in complex with strontium: implication for its secondary effect in bones.
  Protein Sci, 15, 1691-1700.
PDB code: 2glq
11029583 B.Asgeirsson, J.B.Hauksson, and G.H.Gunnarsson (2000).
Dissociation and unfolding of cold-active alkaline phosphatase from atlantic cod in the presence of guanidinium chloride.
  Eur J Biochem, 267, 6403-6412.  
10841753 J.Matysik, A.Alia, G.Nachtegaal, H.J.van Gorkom, A.J.Hoff, and Groot (2000).
Exploring the calcium-binding site in photosystem II membranes by solid-state (113)Cd NMR.
  Biochemistry, 39, 6751-6755.  
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.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.