Structural and functional similarities between osmotin from Nicotiana tabacum seeds and human adiponectin

Marco Miele, Susan Costantini, Giovanni Colonna, Marco Miele, Susan Costantini, Giovanni Colonna

Abstract

Osmotin, a plant protein, specifically binds a seven transmembrane domain receptor-like protein to exert its biological activity via a RAS2/cAMP signaling pathway. The receptor protein is encoded in the gene ORE20/PHO36 and the mammalian homolog of PHO36 is a receptor for the human hormone adiponectin (ADIPOR1). Moreover it is known that the osmotin domain I can be overlapped to the β-barrel domain of adiponectin. Therefore, these observations and some already existing structural and biological data open a window on a possible use of the osmotin or of its derivative as adiponectin agonist. We have modelled the three-dimensional structure of the adiponectin trimer (ADIPOQ), and two ADIPOR1 and PHO36 receptors. Moreover, we have also modelled the following complexes: ADIPOQ/ADIPOR1, osmotin/PHO36 and osmotin/ADIPOR1. We have then shown the structural determinants of these interactions and their physico-chemical features and analyzed the related interaction residues involved in the formation of the complexes. The stability of the modelled structures and their complexes was always evaluated and controlled by molecular dynamics. On the basis of these results a 9 residues osmotin peptide was selected and its interaction with ADIPOR1 and PHO36 was modelled and analysed in term of energetic stability by molecular dynamics. To confirm in vivo the molecular modelling data, osmotin has been purified from nicotiana tabacum seeds and its nine residues peptide synthesized. We have used cultured human synovial fibroblasts that respond to adiponectin by increasing the expression of IL-6, TNF-alpha and IL-1beta via ADIPOR1. The biological effect on fibroblasts of osmotin and its peptide derivative has been found similar to that of adiponectin confirming the results found in silico.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Alignment of adiponectin monomer A…
Figure 1. Alignment of adiponectin monomer A in mouse and human.
Secondary structure prediction made by JPred. Amino acids in the beta-strands are underlined.
Figure 2. 3D model of monomer A…
Figure 2. 3D model of monomer A and trimer of human adiponectin.
The beta-strands are indicated with arrows.
Figure 3. Alignment of Rhodopsin, ADIPOR1 and…
Figure 3. Alignment of Rhodopsin, ADIPOR1 and PHO36.
Secondary structure predictions made by JPred are reported for ADIPOR1 and PHO36. Amino acids in the helices are reported in grey but those interacting with the ADIPOQ, osmotin (OSM) and PeptideOSM in the related complexes with ADIPOR1 and PHO36are evidenced in bold.
Figure 4. 3D model of three complexes.
Figure 4. 3D model of three complexes.
(A) AdipoQ/ADIPOR1 complex, (B) Osmotin/ADIPOR1 complex and (C) PeptideOSM/ADIPOR1 complex with a zoom on the PeptideOSM structure.
Figure 5. Details of Pept OSM -…
Figure 5. Details of PeptOSM - ADIPOR1 interaction.
The peptide is reported by stick representation and yellow labels. The interaction residues of ADIPOR1 are shown by lines and white labels.
Figure 6. SDS-PAGE analysis of osmotin purified…
Figure 6. SDS-PAGE analysis of osmotin purified according to Shih et al. (2001) .
A. Molecular mass marker (LMW, Amersham Pharmacia Biotech); B. Protein is indicated by arrow. The gels were stained with Coomassie brilliant blue.

References

    1. van Loon LC, Rep M, Pieterse CMJ. Significance of Inducible Defense-related Proteins in Infected Plants. Annu Rev Phytopathol. 2006;44:135–62.
    1. Narashiman ML, Coca MA, Jin J, Yamauchi T, Ito Y, et al. Osmotin is a homolog of mammalian adiponectin and controls apoptosis in yeast through a homolog of mammalian adiponectin receptor. Mol Cell. 2005;17:171–180.
    1. Narasimhan ML, Damsz B, Coca MA, Ibeas JI, Yun DJ, et al. A plant defense response effector induces microbial apoptosis. Molecular Cell, 2001;8:921–930.
    1. Yun D-J, Ibeas JI, Lee H, Coca MA, Narasimhan ML, et al. Osmotin, a Plant Antifungal Protein, Subverts Signal Transduction to Enhance Fungal Cell Susceptibility. Molecular Cell. 1998;1:807–817.
    1. Min K, Ha SC, Hasegawa PM, Bressan RA, Yun D-J, et al. Crystal Structure of Osmotin, a Plant Antifungal Protein. Proteins. 2004;54:170–173.
    1. Thompson JD, Higgins DG, Gibson T. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994;22:4673–4680.
    1. Sali A, Blundell TL. Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol. 1993;234:779–815.
    1. Eswar N, Eramian D, Webb B, Shen MY, Sali A. Protein structure modeling with MODELLER. Methods Mol Biol. 2008;426:145–159.
    1. Sippl MJ. Recognition of errors in three-dimensional structures of proteins. Proteins. 1993;17:355–362.
    1. Laskowski RA, MacArthur MW, Moss DS, Thornton JM. PROCHECK: a program to check the stereochemical quality of protein structures. J Appl Cryst. 1993;26:283–291.
    1. Xiang Z, Soto CS, Honig B. Evaluating conformational free energies: the colony energy and its application to the problem of loop prediction. PNAS. 2002;99:7432–7437.
    1. Kabsch W, Sander C. Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features. Biopolymers. 1983;2:2577–2637.
    1. Orengo CA, Michie AD, Jones S, Jones DT, Swindells MB, et al. CATH – a hierarchic classification of protein domain structures. Structure. 1997;5:1093–1108.
    1. Cuff JA, Barton GJ. Application of enhanced multiple sequence alignment profiles to improve protein secondary structure prediction. Proteins. 2000;40:502–511.
    1. Costantini S, Rossi M, Colonna G, Facchiano AM. Modelling of HLA-DQ2 and its interaction with gluten peptides to explain molecular recognition in celiac disease. J Mol Graph Model. 2005;23:419–431.
    1. Costantini S, Colonna G, Facchiano AM. Simulation of conformational changes occurring when a protein interacts with its receptor. Comp Biol Chem. 2007;31:196–206.
    1. Costantini S, Buonocore F, Facchiano AM. Molecular modelling of co-receptor CD8αα and its complex with MHC class I and T-cell receptor in sea bream (Sparus aurata). Fish Shellfish Immunology. 2008;25:782–790.
    1. Gianfrani C, Siciliano R, Facchiano AM, Camarca A, Mazzeo FM, et al. Transamidation of wheat flour inhibits the response to gliadin of intestinal T cells in celiac disease. Gastroenterology. 2007;133:780–789.
    1. Paladino A, Costantini S, Colonna G, Facchiano AM. Molecular modelling of miraculin: structural analyses and functional hypotheses. BBRC. 2008;367:26–32.
    1. Jones S, Thornton JM. Principles of protein-protein interactions derived from structural studies. PNAS. 1996;93:13–20.
    1. Hubbard SJ, Campbell SF, Thornton JM. Molecular recognition. Conformational analysis of limited proteolytic sites and serine proteinase protein inhibitors. J Mol Biol. 1991;220:507–530.
    1. McDonald IK, Thornton JM. Satisfying hydrogen bonding potential in proteins. J Mol Biol. 1994;238:777–793.
    1. Costantini S, Colonna G, Facchiano AM. ESBRI: a software for evaluating the salt bridges in proteins. Bioinformation. 2008;3:137–139.
    1. Autiero I, Costantini S, Colonna G. Human sirt-1: molecular modeling and structure-function relationships of an unordered protein. PLoS One. 2009;4:e7350.
    1. Teodorescu O, Galor T, Pillardy J, Elber R. Enriching the sequence substitution matrix by structural information. Proteins. 2004;54:41–48.
    1. Dalton JAR, Jackson RM. An evaluation of automated homology modelling methods at low target-template sequence similarity. Bioinformatics. 2007;23:1901–1908.
    1. Schneidman-Duhovny D, Inbar Y, Polak V, Shatsky M, Halperin I, et al. Taking geometry to its edge: fast unbound rigid (and hinge-bent) docking. Proteins. 2003;52:107–112.
    1. Schneidman-Duhovny D, Inbar Y, Nussinov R, Wolfson HJ. PatchDock and SymmDock: servers for rigid and symmetric docking. Nucleic Acids Res. 2005;33:W363–W367.
    1. Mashiach E, Nussinov R, Wolfson HJ. FiberDock: a web server for flexible induced-fit backbone refinement in molecular docking. Nuclei Acids Res. 2010;38:W457–W461.
    1. Mashiach E, Schneidman-Duhovny D, Peri A, Shavit Y, Nussinov R, et al. An integrated suite of fast docking algorithms. Proteins. 2010;78:3197–3204.
    1. Van Der Spoel D, Lindahl E, Hess B, Groenhof G, Mark AE, et al. GROMACS: fast, flexible, and free. J Comput Chem. 2005;26:1701–1718.
    1. Costantini S, Facchiano AM. Human Prion Protein Helices: Studying their Stability by Molecular Dynamics Simulations. Protein Pept Lett. 2009;16:1057–1062.
    1. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T. How to measure and predict the molar absorption coefficient of a protein. Protein Science. 1995;4:2411–2423.
    1. Shih C-YT, Wu J, Jia S, Khan AA, Ting K-LH, et al. Purification of an osmotin-like protein from the seeds of Benincasa hispida and cloning of the gene encoding this protein. Plant Science. 2001;160:817–826.
    1. Massimo Mariconda M, Cozzolino A, Attingenti P, Cozzolino F, Milano C. Osteoarticular tuberculosis in a developed country. Journal of Infection. 2007;54:375-e380.
    1. Tang C-H, Chiu YC, Tan TW, Yang RS, Fu W-M. Adiponectin enhances IL-6 production in human synovial fibroblast via an AdipoR1 receptor, AMPK, p38, and NF-kappa B pathway. Journal of Immunology. 2007;179:5483–5492.
    1. Capone F, Costantini S, Guerriero E, Calemma R, Napolitano M, et al. Serum cytokine levels in patients with hepatocellular carcinoma. Eur Cyt Net. 2010;21:99–104.
    1. Yamauchi T, Kamon J, Ito Y, Tsuchida A, Yokomizo T, et al. Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature. 2003;423:762–769.
    1. Kadowaki T, Yamauchi T. Adiponectin and adiponectin receptors. Endocr Rev. 2005;26:439–451.
    1. Liu S, Zhang C, Zhou H, Zhou Y. A physical reference state unifies the structure-derived potential of mean force for protein folding and binding. Proteins. 2004;56:93–101.
    1. Costantini S, Miele M, Colonna G. Intrinsically Unordered Proteins: structural properties, prediction and relevance in Protein Folding, Chapter 6 - Editors: Eric C. Walters Series: Protein Biochemistry, Synthesis, Structure and Cellular Functions, Nova Science Publishers, Inc. Hauppauge NY – USA ISBN. 2010:978–1–61761–259–6.
    1. Trotta T, Costantini S, Colonna G. Modelling of the membrane receptor CXCR3 and its complexes with CXCL9, CXCL10 and CXCL11 chemokines: putative target for new drug design. Molecular Immunology. 2009;47:332–339.

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