PDBsum entry 3dpc

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Hydrolase PDB id
Protein chains
449 a.a. *
Waters ×221
* Residue conservation analysis
PDB id:
Name: Hydrolase
Title: Structure of e.Coli alkaline phosphatase mutant in complex w phosphorylated peptide
Structure: Alkaline phosphatase. Chain: a, b. Synonym: apase. Engineered: yes. Mutation: yes. Phosphorylated peptide. Chain: c. Engineered: yes
Source: Escherichia coli. Organism_taxid: 83333. Strain: k12. Gene: phoa, b0383, jw0374. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Other_details: this sequence is chemically synthesized
2.30Å     R-factor:   0.234     R-free:   0.271
Authors: W.H.Wang,T.Jiang
Key ref: W.Li et al. (2009). Development of a universal phosphorylated peptide-binding protein for simultaneous assay of kinases. Biosens Bioelectron, 24, 2871-2877. PubMed id: 19349157
07-Jul-08     Release date:   16-Jun-09    
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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 1 residue position (black cross)

 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
Bound ligand (Het Group name = TPO)
matches with 45.45% similarity
      Cofactor: Mg(2+); Zn(2+)
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  


Biosens Bioelectron 24:2871-2877 (2009)
PubMed id: 19349157  
Development of a universal phosphorylated peptide-binding protein for simultaneous assay of kinases.
W.Li, L.Bi, W.Wang, Y.Li, Y.Zhou, H.Wei, T.Jiang, L.Bai, Y.Chen, Z.Zhang, X.Yuan, J.Xiao, X.E.Zhang.
This study describes the development of a universal phosphorylated peptide-binding protein designed to simultaneously detect serine, threonine and tyrosine kinases. The Escherichia coli alkaline phosphatase (EAP) is a well-defined nonspecific phosphated monoesterase and Ser-, Thr- or Tyr-phosphorylated peptides served as substrates for EAP in preliminary experiments. Based on the known catalytic mechanism of EAP, the recombinant site-directed mutant EAP-S102L was generated, whose catalytic activity was blocked, but its binding ability was preserved. For EAP-S102L the catalytic rate constant, k(cat), was reduced by a factor of 1000, while the Michaelis-Menten constant, K(m), remained almost unchanged. Crystallographic analysis of the EAP-S102L/phophorylated peptide complex revealed that EAP-S102L could bind the phosphate group of the phosphorylated peptide but lacked nucleophilic attack potential which was essential for the catalytic ability of EAP. Finally, by combining the fluorescence-labeled EAP-S102L with non-phophorylated peptide chips, kinases could be detected from tumor cell samples. The recombinant EAP-S102L construct is perhaps the first functional binding protein derived from a native enzyme, illustrating how one single mutation tremendously alters protein function.