PDBsum entry 6drb

Hydrolase

PDB id: 6drb

Contents

Protein chain

278 a.a.

Ligands

WO4

PDB id:

6drb

Name:

Hydrolase

Title:

Crystal structure of the yoph ptp1b wpd loop chimera 3 ptpase bound to tungstate

Structure:

Targeted effector protein. Chain: a. Synonym: type iii secretion injected virulence protein,yoph. Engineered: yes. Mutation: yes

Source:

Yersinia pestis. Organism_taxid: 632. Gene: yoph, ypcd1.67c. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

2.75Å R-factor: 0.215 R-free: 0.264

Authors:

Y.Morales,S.J.Johnson,A.C.Hengge

Key ref:

G.Moise et al. (2018). A YopH PTP1B Chimera Shows the Importance of the WPD-Loop Sequence to the Activity, Structure, and Dynamics of Protein Tyrosine Phosphatases. Biochemistry, 57, 5315-5326. PubMed id: 30110154 DOI: 10.1021/acs.biochem.8b00663

Date:

11-Jun-18 Release date: 29-Aug-18

Protein chain

O68720  (O68720_YERPE) -  protein-tyrosine-phosphatase from Yersinia pestis

Seq: 468 a.a.

Struc: 278 a.a.*

* PDB and UniProt seqs differ at 5 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

DOI no: 10.1021/acs.biochem.8b00663

Biochemistry 57:5315-5326 (2018)

PubMed id: 30110154

A YopH PTP1B Chimera Shows the Importance of the WPD-Loop Sequence to the Activity, Structure, and Dynamics of Protein Tyrosine Phosphatases.

G.Moise, Y.Morales, V.Beaumont, T.Caradonna, J.P.Loria, S.J.Johnson, A.C.Hengge.

ABSTRACT

To study factors that affect WPD-loop motion in protein tyrosine phosphatases (PTPs), a chimera of PTP1B and YopH was created by transposing the WPD loop from PTP1B to YopH. Several subsequent mutations proved to be necessary to obtain a soluble, active enzyme. That chimera, termed chimera 3, retains productive WPD-loop motions and general acid catalysis with a pH dependency similar to that of the native enzymes. Kinetic isotope effects show the mechanism and transition state for phosphoryl transfer are unaltered. Catalysis of the chimera is slower than that of either of its parent enzymes, although its rate is comparable to those of most native PTPs. X-ray crystallography and nuclear magnetic resonance were used to probe the structure and dynamics of chimera 3. The chimera's structure was found to sample an unproductive hyper-open conformation of its WPD loop, a geometry that has not been observed in either of the parents or in other native PTPs. The reduced catalytic rate is attributed to the protein's sampling of this conformation in solution, reducing the fraction in the catalytically productive loop-closed conformation.