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PDBsum entry 2fmj
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protein ligands metals links
Hydrolase PDB id
2fmj

 

 

 

 

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Contents
Protein chain
222 a.a. *
Ligands
SO4
Metals
_CA
Waters ×272
* Residue conservation analysis
PDB id:
2fmj
Name: Hydrolase
Title: 220-loop mutant of streptomyces griseus trypsin
Structure: Trypsin. Chain: a. Synonym: sgt. Engineered: yes. Mutation: yes
Source: Streptomyces griseus. Organism_taxid: 1911. Gene: sprt. Expressed in: bacillus subtilis. Expression_system_taxid: 1423.
Resolution:
1.65Å     R-factor:   0.157     R-free:   0.189
Authors: M.J.Page,E.Di Cera
Key ref:
M.J.Page et al. (2006). Conversion of trypsin into a Na(+)-activated enzyme. Biochemistry, 45, 2987-2993. PubMed id: 16503653 DOI: 10.1021/bi052481a
Date:
09-Jan-06     Release date:   23-May-06    
PROCHECK
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 Headers
 References

Protein chain
Pfam   ArchSchema ?
P00775  (TRYP_STRGR) -  Trypsin from Streptomyces griseus
Seq:
Struc: 		259 a.a.
222 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 4 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.3.4.21.4  - trypsin.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Preferential cleavage: Arg-|-Xaa, Lys-|-Xaa.

 

 
DOI no: 10.1021/bi052481a Biochemistry 45:2987-2993 (2006)
PubMed id: 16503653  
 
 
Conversion of trypsin into a Na(+)-activated enzyme.
M.J.Page, M.R.Bleackley, S.Wong, R.T.MacGillivray, E.Di Cera.
 
  ABSTRACT  
 
Serine proteases of the chymotrypsin family show a dichotomous amino acid distribution for residue 225. Enzymes carrying Tyr at position 225 are activated by Na(+), whereas those carrying Pro are devoid of Na(+) binding and activation. Previous studies have demonstrated that the Y225P conversion is sufficient to abrogate Na(+) activation in several enzymes. However, the reverse substitution P225Y is necessary but not sufficient to introduce Na(+) binding and activation. Here we report that Streptomyces griseus trypsin, carrying Pro-225, can be engineered into a Na(+)-activated enzyme by replacing residues in the 170, 186, and 220 loops to those of coagulation factor Xa. The findings represent the first instance of an engineered Na(+)-activated enzyme and a proof of principle that should enable the design of other proteases with enhanced catalytic activity and allosteric regulation mediated by monovalent cation binding.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
18377928 M.J.Page, C.J.Carrell, and E.Di Cera (2008).
Engineering protein allostery: 1.05 A resolution structure and enzymatic properties of a Na+-activated trypsin.
  J Mol Biol, 378, 666-672.
PDB code: 3beu
17347701 E.Di Cera, M.J.Page, A.Bah, L.A.Bush-Pelc, and L.C.Garvey (2007).
Thrombin allostery.
  Phys Chem Chem Phys, 9, 1291-1306.  
17428793 F.Marino, Z.W.Chen, C.E.Ergenekan, L.A.Bush-Pelc, F.S.Mathews, and E.Di Cera (2007).
Structural basis of Na+ activation mimicry in murine thrombin.
  J Biol Chem, 282, 16355-16361.
PDB codes: 2ocv 2od3
17636263 L.A.Bush-Pelc, F.Marino, Z.Chen, A.O.Pineda, F.S.Mathews, and E.Di Cera (2007).
Important role of the cys-191 cys-220 disulfide bond in thrombin function and allostery.
  J Biol Chem, 282, 27165-27170.
PDB codes: 2pgb 2pgq
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.

 

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