PDBsum entry 5tnh

Hydrolase

PDB id: 5tnh

Contents

Protein chains

293 a.a.

Ligands

7MS ×4

Waters ×600

PDB id:

5tnh

Name:

Hydrolase

Title:

Crystal structure of the e153q mutant of the cftr inhibitory factor cif containing the adducted 17,18-epete hydrolysis intermediate

Structure:

Cftr inhibitory factor. Chain: c, d, a, b. Engineered: yes. Mutation: yes

Source:

Pseudomonas aeruginosa (strain ucbpp-pa14). Organism_taxid: 208963. Strain: ucbpp-pa14. Gene: pa14_26090. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.10Å R-factor: 0.188 R-free: 0.234

Authors:

K.L.Hvorecny,D.R.Madden

Key ref:

K.L.Hvorecny et al. (2017). Active-Site Flexibility and Substrate Specificity in a Bacterial Virulence Factor: Crystallographic Snapshots of an Epoxide Hydrolase. Structure, 25, 697. PubMed id: 28392259

Date:

14-Oct-16 Release date: 11-Oct-17

Protein chains

A0A0H2ZD27  (A0A0H2ZD27_PSEAB) -  Putative hydrolase from Pseudomonas aeruginosa (strain UCBPP-PA14)

Seq: 319 a.a.

Struc: 293 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Structure 25:697 (2017)

PubMed id: 28392259

Active-Site Flexibility and Substrate Specificity in a Bacterial Virulence Factor: Crystallographic Snapshots of an Epoxide Hydrolase.

K.L.Hvorecny, C.D.Bahl, S.Kitamura, K.S.S.Lee, B.D.Hammock, C.Morisseau, D.R.Madden.

ABSTRACT

Pseudomonas aeruginosa secretes an epoxide hydrolase with catalytic activity that triggers degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) and perturbs other host defense networks. Targets of this CFTR inhibitory factor (Cif) are largely unknown, but include an epoxy-fatty acid. In this class of signaling molecules, chirality can be an important determinant of physiological output and potency. Here we explore the active-site chemistry of this two-step α/β-hydrolase and its implications for an emerging class of virulence enzymes. In combination with hydrolysis data, crystal structures of 15 trapped hydroxyalkyl-enzyme intermediates reveal the stereochemical basis of Cif's substrate specificity, as well as its regioisomeric and enantiomeric preferences. The structures also reveal distinct sets of conformational changes that enable the active site to expand dramatically in two directions, accommodating a surprising array of potential physiological epoxide targets. These new substrates may contribute to Cif's diverse effects in vivo, and thus to the success of P. aeruginosa and other pathogens during infection.