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Characterization of the adipogenic protein E4orf1 from adenovirus 36 through an in silico approach

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Abstract

Adenovirus 36 (Ad-36) is related to human obesity due to its adipogenic activity mediated by the early 4 open reading frame 1 (E4orf1) protein. Mechanisms underlying the adipogenic effect of E4orf1 are not completely understood; however, the proliferation and differentiation of fat cells are increased through the activation of the phosphatidyl inositol 3 kinase pathway by binding proteins containing PDZ domain. This study characterized E4orf1 tridimensional structure and analyzed its interactions with PDZ domain-containing proteins in order to provide new information about the behavior of this viral protein and its targets, which could provide an interesting druggable target for obesity-related cardiometabolic alterations. In silico strategies such as homology modeling, docking, and molecular dynamics (MD) were used to study the interaction of E4orf1 with five PDZ domains of disk large homolog 1 (PDZ-1 and PDZ-2), membrane-associated guanylate kinase 1 (PDZ-3), and multi-PDZ domain protein 1 (PDZ-7 and PDZ-10). Mutagenesis analysis of selected residues was performed to evaluate their effects on the stabilization of E4orf1:PDZ complexes. MD simulations showed that the E4orf1:PDZ10 complex was more stable than the others ones. The highly hydrophobic residues at the C-terminal region (114–125) of the E4orf1 are essential in the initial phase stabilization of the complexes. Moreover, the residues 80–85 in the core region contribute to longer stabilization of the E4orf1:PDZ10 complex, a result that was confirmed by in silico mutagenesis. In conclusion, E4orf1 forms a stable complex with PDZ10 domain, and the residues 80–85 are of particular importance. The characterization of E4orf1 interactions with PDZ domains provides an initial approach to discover druggable targets for Ad-36-induced obesity.

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Abbreviations

Ad-36:

Adenovirus 36

cAMP:

Cyclic adenosine monophosphate

DLG1:

Disk large homolog 1

DOPE:

Discrete optimized protein energy

E4orf1:

Early 4 open reading frame 1

PI3K:

Phosphatidyl inositol 3 kinase

MAGI-1:

Membrane-associated guanylate kinase 1

MD:

Molecular dynamics

MUPP1:

Multi-PDZ domain protein 1

PATJ:

PALS1-associated tight junction protein

PBM:

PDZ-binding motif

ZO-2:

Zona occludens 2

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Acknowledgments

Graphics were obtained from GraphPad Prims v.7. Pictures of models and atoms interactions were obtained from PyMOL v2.3. All images were edited with Inkscape 0.92.4.

Funding

This study was supported by Fondecyt-Chile (Grant number #11150445) and PCI-Conicyt (Grant number #REDI170632). We thank the Centro de Modelación y Computación Cientifica at the Universidad de la Frontera (CMCC-UFRO) and the Laboratory of Advanced Scientific Computing at the University of Sao Paulo by providing the supercomputing infrastructure where MD was performed. MH Hirata and RDC Hirata are recipients of fellowships from CNPq. GM Ferreira is a fellowship recipient from FAPESP-Brazil (Grant number #2019/06172-4).

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Author notes

  1. Alvaro Gutiérrez and Glaucio Monteiro Ferreira contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Bioanalysis and Molecular Diagnosis, Center of Excellence in Translational Medicine, CEMT-BIOREN, Universidad de La Frontera, Temuco, Chile

    Alvaro Gutiérrez & Alvaro Cerda

  2. Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo, Brazil

    Glaucio Monteiro Ferreira, Mario Hirouyki Hirata & Rosario Dominguez Crespo Hirata

  3. Laboratory of Molecular Investigation in Cardiology, Dante Pazzanese Institute of Cardiology, Sao Paulo, Brazil

    Glaucio Monteiro Ferreira, Fausto Feres, Mario Hirouyki Hirata & Rosario Dominguez Crespo Hirata

  4. Laboratory of Ecological Chemistry, Department of Natural Resource and Chemical Science, Universidad de La Frontera, Temuco, Chile

    Juan Machuca & Herbert Venthur

  5. Department of Basic Sciences, Universidad de La Frontera, Temuco, Chile

    Alvaro Cerda

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  1. Alvaro Gutiérrez
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Gutiérrez, A., Ferreira, G.M., Machuca, J. et al. Characterization of the adipogenic protein E4orf1 from adenovirus 36 through an in silico approach. J Mol Model 26, 285 (2020). https://doi.org/10.1007/s00894-020-04531-0

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