# Structural view of fungal glutathionyl-hydroquinone reductases

C. Didierjean, Mathieu Schwartz, Thomas Perrot, Arnaud Hecker, Éric

Gelhaye, Frederique Favier

► To cite this version:

C. Didierjean, Mathieu Schwartz, Thomas Perrot, Arnaud Hecker, Éric Gelhaye, et al. Structural view of fungal glutathionyl-hydroquinone reductases. 30. European Crystallographic Meeting, Aug 2016, Basel, Switzerland. Wiley-Blackwell, Acta crystallographica. Section A, Foundations of crystallography, A72, 1 p., 2016, 10.1107/S205327331609656X. http://hal-02743022

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Distributed under a Creative Commons Attribution 4.0 International License
MS9-P2 Structural view of fungal
glutathionyl-hydroquinone reductases

Claude DIDIERJEAN¹,², Mathieu Schwartz¹, Thomas Perrot²,
Arnaud Hecker², Eric Gelhaye³, Frédéric Favier¹

1. University of Lorraine, UMR 7036 CRM2,
Vandoeuvre-les-Nancy, France

2. CNRS, UMR 7036 CRM2, Vandoeuvre-les-Nancy, France

email: claude didierjean@univ-lorraine.fr

Glutathione transferases (GSTs) constitute a widespread
super-family of enzymes with highly specific to
overlapping functions. These enzymes have been
extensively studied in mammals because of their roles in
enoxibiotic conjugation. Despite this, GST functions are
largely unexplored, especially in microorganisms. Our
work focused on fungal glutathionyl-hydroquinone
reductases (GHRs). These singular GSTs catalyze the
reduction of hydroquinones-SG (Xun et al., 2010), and
are present in all kingdoms but animals. The first GHR
structure to be described in the saprotrophic fungus
*Phanerochaete chrysosporium* (PeGHR1) has unveiled
original features. An original dimerization mode led us to
establish a new structural class named Xi (GSTX) (Meux
et al., 2011).

Here we report the crystal structures of two GHRs, TvGHR1 from the basidiomycete *Trametes versicolor* and ScECM4 from the yeast *Saccharomyces cerevisiae*, at 2.28 and 1.45 Å resolution, respectively. TvGHR1 and ScECM4 share the classical GST fold which consists in a thioredoxin N-terminal domain and an all α C-terminal domain. Besides the features that have been uncovered in PeGHR1, additional secondary structures were found in the yeast ScECM4 structure. Like its characterized homologs, ScECM4 is specific to the reduction of quinones conjugated with glutathione (Lallement *et al.*, 2014). TvGHR1 is more polyvalent because it also reduces substrates rather accepted by Omega-class GSTs like phenylacetophenone-SG. Compared with known GHR structures, TvGHR1 exhibits a larger substrate binding site with an original motif that could explain the substrate versatility. Phylogenetic analyses revealed that fungal GHRs could be separated in two families : the 'classic' GHRs and the 'atypical' GHRs that exhibit motif variations as observed in TvGHR1. Furthermore, the conservation of GHRs in fungi suggests that they could fulfill a crucial role. Their diversification in wood-degrading basidiomycetes agrees with the hypothesis that GHRs would play a role in the metabolism of phenolic compounds (Belchik & Xun, 2011).

Belchick S.M. & Xun L., Drug Metab Rev. 2011, 43(2),
307 ; Lallement et al., FEBS Lett. 2015, 589, 37; Meux et
al., J. Biol. Chem. 2011, 286(11), 9162 ; Xun et al.,
Biochem. J. 2010, 428(3), 419.

Keywords: glutathionyl-hydroquinone, reductase, fungi, glutathione transferase

MS9-P3 PLA₂S-like membrane perturbation
mechanism: extracting the most of
crystallography data

Rafael J. Borges¹,², Ney Lemke¹, Massimo Sammito², Claudia Millán², Isabel Usón³, Marcos M.R. Fontes¹

1. Dept. of Physics and Biophysics, Biosciences Institute (IBB),
São Paulo State University (UNESP), Botucatu, São Paulo, Brazil

2. Dept. of Structural Biology, Institute of Molecular Biology of Barcelona (IBMB), Spanish National Research Council (CSIC), Barcelona, Spain

3. Dept. of Structural Biology, Catalan Institution for Research and Advanced Studies (ICREA), Barcelona, Spain

email: rjborges@ibb.unesp.br

Venomics has been an invaluable tool to biomedicine
by aiding comprehension of physiological mechanisms
and drug design. Particularly, secreted phospholipases A₂,
(PLA₂S) had their catalytic mechanism first inferred
evaluating three high resolution crystallographic
structures PLA₂S from Chinese cobra and bee venom in
different states²⁻¹. A subtype of PLA₂S, the PLA₂S-like,
also named as PLA₂S-homologue due to its similar
tertiary structure and common ancestor, does not possess
catalytic activity but still presents a high myotoxicity and
a wide range of pharmacological activities²,³. The
PLA₂S-like versatility has related to its capability to
perturb different biological membrane, breaking ionic
homeostasis and inducing series physiological events that
leads to cell death ⁵. Although a lot has been discovered
by functional and structural experiments, PLA₂S-like
exact mechanism of action is not fully understood ²,³.
Herein, we investigated their molecular mechanism by
evaluation of global and local characteristics of
crystallographic models. We gathered all snake venom
PLA₂S-like structures available in Protein DataBank in
the most probable dimer in solution using ProtCID
database⁴. We evaluated models by hydrophobic channel
accessibility through MOLE 2.0.⁵, by presence or absence
of hydrophobic ligands and by the geometrical orientation
of one monomer to the other. The geometrical orientation
was extracted from the rotation matrix after monomer
superimposition using Isqkab program ⁶ available in CCP4
suite ⁷. By these global and local structural evaluations,
we identified different native and complex states relating
monomer-monomer geometry to hydrophobic channel
aperture and closure.

References

1. Scott, D. L. et al. *Science* **250**, 1541–1546 (1990).

2. Fernandes, C. A. H., et al. Borges, R. J., Lomonte, B. & Fontes, M. R., M. Biochim. Biophys. Acta (2014). doi:10.1016/j.bgapap.2014.09.015
3. Lomonte, B., Angulo, Y., Sasa, M. & Gutiérrez, J. M. *Protein Pept* *Lett.* **16**, 860–876 (2009).
4. Xu, Q. & Dunbrack, R. L. *Nucleic Acids Res.* **39**, D761–770 (2011).
5. Sehnal, D. *et al*. MOLE 2.0: advanced approach for analysis of biomacromolecular channels. *J. Cheminformatics* **5**, 39 (2013).
6. Kabsch, W. *Acta Crystallogr. Sect. A* **32**, 922–923 (1976).
7. Winn, M. D. *et al*. *Acta Crystallogr. D Biol. Crystallogr.* **67**, 235–242 (2011).

Keywords: Phospholipase A2, geometry, hydrophobic channel