PDBsum entry 3f99

Hydrolase

PDB id: 3f99

Contents

Protein chain

282 a.a. *

Ligands

TRS

Waters ×268

* Residue conservation analysis

PDB id:

3f99

Name:

Hydrolase

Title:

W354f yersinia enterocolitica ptpase apo form

Structure:

Tyrosine-protein phosphatase yoph. Chain: a. Fragment: yoph catalytic domain: unp residues 164-468. Synonym: virulence protein. Engineered: yes. Mutation: yes

Source:

Yersinia enterocolitica (type o:9). Organism_taxid: 34055. Strain: w22703 / serotype o:9 / biotype 2. Gene: yoph, yop51. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.65Å R-factor: 0.198 R-free: 0.234

Authors:

T.A.S.Brandao,H.Robinson,S.J.Johnson,A.C.Hengge

Key ref:

T.A.Brandão et al. (2009). Impaired acid catalysis by mutation of a protein loop hinge residue in a YopH mutant revealed by crystal structures. J Am Chem Soc, 131, 778-786. PubMed id: 19140798

Date:

13-Nov-08 Release date: 20-Jan-09

Protein chain

P15273  (YOPH_YEREN) -  Tyrosine-protein phosphatase YopH from Yersinia enterocolitica

Seq: 468 a.a.

Struc: 282 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.1.3.48 - protein-tyrosine-phosphatase.
• Reaction: O-phospho-L-tyrosyl-[protein] + H2O = L-tyrosyl-[protein] + phosphate

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Key reference

J Am Chem Soc 131:778-786 (2009)

PubMed id: 19140798

Impaired acid catalysis by mutation of a protein loop hinge residue in a YopH mutant revealed by crystal structures.

T.A.Brandão, H.Robinson, S.J.Johnson, A.C.Hengge.

ABSTRACT

Catalysis by the Yersinia protein-tyrosine phosphatase YopH is significantly impaired by the mutation of the conserved Trp354 residue to Phe. Though not a catalytic residue, this Trp is a hinge residue in a conserved flexible loop (the WPD-loop) that must close during catalysis. To learn why this seemingly conservative mutation reduces catalysis by 2 orders of magnitude, we have solved high-resolution crystal structures for the W354F YopH in the absence and in the presence of tungstate and vanadate. Oxyanion binding to the P-loop in W354F is analogous to that observed in the native enzyme. However, the WPD-loop in the presence of oxyanions assumes a half-closed conformation, in contrast to the fully closed state observed in structures of the native enzyme. This observation provides an explanation for the impaired general acid catalysis observed in kinetic experiments with Trp mutants. A 1.4 A structure of the W354F mutant obtained in the presence of vanadate reveals an unusual divanadate species with a cyclic [VO](2) core, which has precedent in small molecules but has not been previously reported in a protein crystal structure.