4GSN

Crystal Structure of GSTe2 ZAN/U variant from Anopheles gambiae

PDB DOI: https://doi.org/10.2210/pdb4GSN/pdb

Classification: TRANSFERASE
Organism(s): Anopheles gambiae
Expression System: Escherichia coli BL21
Mutation(s): Yes

Deposited: 2012-08-28 Released: 2014-03-12
Deposition Author(s): Mayans, O., Lu, F.

Experimental Data Snapshot

Method: X-RAY DIFFRACTION
Resolution: 2.30 Å
R-Value Free: 0.225
R-Value Work: 0.173
R-Value Observed: 0.175

wwPDB Validation 3D Report Full Report

Ligand Structure Quality Assessment

This is version 1.3 of the entry. See complete history.

Literature

Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae.

Mitchell, S.N., Rigden, D.J., Dowd, A.J., Lu, F., Wilding, C.S., Weetman, D., Dadzie, S., Jenkins, A.M., Regna, K., Boko, P., Djogbenou, L., Muskavitch, M.A., Ranson, H., Paine, M.J., Mayans, O., Donnelly, M.J.

(2014) PLoS One 9: e92662-e92662

PubMed: 24675797 Search on PubMedSearch on PubMed Central
DOI: https://doi.org/10.1371/journal.pone.0092662
Primary Citation of Related Structures:
4GSN

PubMed Abstract:
The development of resistance to insecticides has become a classic exemplar of evolution occurring within human time scales. In this study we demonstrate how resistance to DDT in the major African malaria vector Anopheles gambiae is a result of both target-site resistance mechanisms that have introgressed between incipient species (the M- and S-molecular forms) and allelic variants in a DDT-detoxifying enzyme. Sequencing of the detoxification enzyme, Gste2, from DDT resistant and susceptible strains of An. gambiae, revealed a non-synonymous polymorphism (I114T), proximal to the DDT binding domain, which segregated with strain phenotype. Recombinant protein expression and DDT metabolism analysis revealed that the proteins from the susceptible strain lost activity at higher DDT concentrations, characteristic of substrate inhibition. The effect of I114T on GSTE2 protein structure was explored through X-ray crystallography. The amino acid exchange in the DDT-resistant strain introduced a hydroxyl group nearby the hydrophobic DDT-binding region. The exchange does not result in structural alterations but is predicted to facilitate local dynamics and enzyme activity. Expression of both wild-type and 114T alleles the allele in Drosophila conferred an increase in DDT tolerance. The 114T mutation was significantly associated with DDT resistance in wild caught M-form populations and acts in concert with target-site mutations in the voltage gated sodium channel (Vgsc-1575Y and Vgsc-1014F) to confer extreme levels of DDT resistance in wild caught An. gambiae.

Organizational Affiliation

Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.

Macromolecule Content

Total Structure Weight: 100.56 kDa
Atom Count: 7,260
Modelled Residue Count: 876
Deposited Residue Count: 880
Unique protein chains: 1

Macromolecules

Find similar proteins by:

(by identity cutoff) | 3D Structure

Entity ID: 1
Molecule	Chains	Sequence Length	Organism	Details	Image
Glutathione S-transferase E2	A, B, C, D	220	Anopheles gambiae	Mutation(s): 3 EC: 2.5.1.18
UniProt
Find proteins for Q9GPL8 (Anopheles gambiae) Explore Q9GPL8 Go to UniProtKB: Q9GPL8
Entity Groups
Sequence Clusters	30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt Group	Q9GPL8
Sequence Annotations Expand
Reference Sequence

Small Molecules

Ligands 3 Unique
ID	Chains	Name / Formula / InChI Key	2D Diagram	3D Interactions
GSH Query on GSH Download Ideal Coordinates CCD File SDF format, chain E [auth A] SDF format, chain G [auth B] SDF format, chain I [auth C] SDF format, chain J [auth D] MOL2 format, chain E [auth A] MOL2 format, chain G [auth B] MOL2 format, chain I [auth C] MOL2 format, chain J [auth D] mmCIF format, chain E [auth A] mmCIF format, chain G [auth B] mmCIF format, chain I [auth C] mmCIF format, chain J [auth D]	E [auth A], G [auth B], I [auth C], J [auth D]	GLUTATHIONE C₁₀ H₁₇ N₃ O₆ S RWSXRVCMGQZWBV-WDSKDSINSA-N		Interactions Focus chain E [auth A] Focus chain G [auth B] Focus chain I [auth C] Focus chain J [auth D] Interactions & Density Focus chain E [auth A] Focus chain G [auth B] Focus chain I [auth C] Focus chain J [auth D]
1PE Query on 1PE Download Ideal Coordinates CCD File SDF format, chain F [auth A] MOL2 format, chain F [auth A] mmCIF format, chain F [auth A]	F [auth A]	PENTAETHYLENE GLYCOL C₁₀ H₂₂ O₆ JLFNLZLINWHATN-UHFFFAOYSA-N		Interactions Focus chain F [auth A] Interactions & Density Focus chain F [auth A]
GOL Query on GOL Download Ideal Coordinates CCD File SDF format, chain H [auth B] MOL2 format, chain H [auth B] mmCIF format, chain H [auth B]	H [auth B]	GLYCEROL C₃ H₈ O₃ PEDCQBHIVMGVHV-UHFFFAOYSA-N		Interactions Focus chain H [auth B] Interactions & Density Focus chain H [auth B]

Experimental Data & Validation

Experimental Data

Method: X-RAY DIFFRACTION
Resolution: 2.30 Å
R-Value Free: 0.225
R-Value Work: 0.173
R-Value Observed: 0.175
Space Group: P 1 2₁ 1

Unit Cell:

Length ( Å )	Angle ( ˚ )
a = 51.33	α = 90
b = 86.38	β = 90.73
c = 92.85	γ = 90

Software Package:

Software Name	Purpose
PHENIX	refinement
PHASER	phasing
REFMAC	refinement
XDS	data reduction
XSCALE	data scaling
DNA	data collection

Structure Validation

View Full Validation Report

Ligand Structure Quality Assessment

Entry History

Deposition Data

Released Date: 2014-03-12

Deposition Author(s):

Mayans, O.

Lu, F.

Revision History (Full details and data files)

Version 1.0: 2014-03-12
Type: Initial release
Version 1.1: 2014-05-07
Changes: Database references
Version 1.2: 2019-07-17
Changes: Data collection, Refinement description
Version 1.3: 2024-02-28
Changes: Data collection, Database references, Derived calculations