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Institut de Biologie StructuraleGrenoble / France

Contact person(s) related to this article / FIESCHI Franck

Membrane and Immunity Team

Team leader: Franck Fieschi (Professor, UGA)

Team

- Eric Forest (DR, CEA).
- Corinne Vivès (Engineer, CEA).
- Jérôme Dupuy (Assistant Professor, UGA).
- Michel Thépaut (Research engineer, CNRS).
- Isabelle Hartlein (Engineer assistant, CNRS).
- Pascal Gabard (Technicien, Industrie).
- Vanessa Porkolab (PhD, MENRT).
- Silvia Achilli (PhD, ITN Marie Curie).
- Nina Breteau (Professional internship).

Introduction

Many steps of cell/cell, host/pathogen interactions and cellular activation are required to access to the immune response. Those events require a broad range of membrane proteins that are often specific of a peculiar cell type. Those proteins of primary relevance to human health (pathogen receptors, chemokine receptors, adhesion molecules) constitute challenging projects for academic research but also for pharmaceutical applications. Despite the difficulties linked to that type of project, considering the methodological pitfalls in eukaryotic membrane protein research, we have initiated three scientific projects in this context:

NADPH oxidase complex: a membrane multi component enzymatic complex

- Contact: Franck FIESCHI.
- Involved staff: Corinne Vivès, Jérôme Dupuy, Michel Thépaut, Isabelle Hartlein, Pascal Gabard, Nina Breteau.

During a bacterial infection, the neutrophil NADPH oxidase initiates the destruction of the phagocyted microorganisms at the infection site by the production of large amounts of superoxide ions and derived products (H2O2, HOCl). The toxicity of the produced species requires a tight regulation of this system by the cell. This macromolecular complex comprises a membrane component the b558 flavocytochrome and several cytosolic proteins. Consecutively to an activation signal the complex associates at the membrane and initiates the electron transfer from NADPH to O2.

We investigate with a structural and functional approach the various components of the system as well as some of the steps leading to the structural modifications involved in the complex assembly at the membrane and to the activation of the enzymatic reaction.

Collaborations:

- IBBMC, Université de Paris XI, Orsay.
- Eric Forest, Institut de Biologie Structurale, Grenoble.
- CDiReC, CHU, Grenoble.
- Tel Aviv University, Israel.

Host-pathogen interactions

- Contact GPCR: Corinne Vivès and Franck Fieschi.
- Involved staff GPCR: Franck Fieschi, Eva Pebay-Peyroula.
- Contact Lectines: Franck Fieschi.
- Involved staff Lectines: Michel Thépaut, Corinne Vivès, Vanessa Porkolab, Silvia Achilli.

During HIV infection, the virus is first captured by the muquous epithelia and then is transported to the lymphoid organs where it reaches its major cellular targets: the T lymphocytes. At the level of the epithelia, the virus is captured via the interaction of its surface glycoproteins with a membrane lectin expressed on the dendritic cell surface: DC-SIGN. Once attached, the intact virus is driven by these dendritic cells to the T lymphocytes. The virus can then infect the target cell through the interaction with several cell receptors including CXCR4 or CCR5. These various proteins are involved in the virus capture, transport and infection and constitute the subject of either methodological developments or of structural and functional studies.

A significant effort has been placed in the development of an overexpression strategy for these G protein coupled receptors, CXCR4 and CCR5. As for DC-SIGN, we characterize the biological and structural properties of this oligomeric receptor and investigate the properties of its interaction with its targets. Collaborations involving several groups from the Institut Pasteur, Paris, on the DC-SIGN project allowed the study of other pathologies in which DC-SIGN has been shown to be involved, including the Dengue, virus C hepatitis or the Cytomegalovirus. Finally, a collaboration with a saccharide chemist group in Spain enables the development of multivalent molecules (glycodendrimers) specific to DC-SIGN in order to inhibit the virus/DC-SIGN interaction or to develop functionalised vectors to target the dendritic cells.

Other lectins from dendritic cells present interaction properties with pathogens. Among these lectins, we work also on Langerine. This specific lectin of Langerhans cells is also able to recognize HIV. Moreover, it is involved in the formation of an original organite, the Birbeck granule. We are studying the physiologic ligands of this lectin, its structural properties as well as its role in Birbeck granule formation.

Collaborations:

- Hugues Lortat–Jacob, LEM Institut de Biologie Structurale, Grenoble.
- Anne Imberty, CERMAV, Grenoble.
- Fernando Arenzana-Seisdedos, Institut Pasteur, Paris.
- Patrice Vachette & Dominique Durand, Université de Paris XI, Orsay.
- Jean-Luc Popot, IBPC, Paris.
- Jean-Louis Bannère, Faculté de Pharmacie, Montpellier.
- Jenny Valladeau, University Claude Bernard, Lyon.
- Javier Rojo, Instituto de Investigaciones Quimicas, Sevilla, Spain.
- Anna Bernardi, Universita’ di Milano, Italy.

Related platform

The MP3 platform (Membrane Protein Purification Platform) corresponds to four protein purification modules of the Akta Xpress type, dedicated to membrane protein purification.

Person in charge of the platform : Michel Thépaut.

Team members:

Back row (left to right): Eric Forest, Franck Fieschi, Jérôme Dupuy, Michel Thépaut, Isabelle Petit-Hartlein.
Front row (left to right): Nina Breteau, Silvia Achilli, Vanessa Porkolab, Pascal Gabard, Corinne Vivès.

Patent

Dupuy J, Hajjar C, Cherrier M, Fieschi F.
NADPH oxidase proteins.
WO/2015/162383 (2015).

Recent publications

Guzzi C, Alfarano P, Sutkeviciute I, Sattin S, Ribeiro-Viana R, Fieschi F, Bernardi A, Weiser J, Rojo J, Angulo J, Nieto PM.
Detection and quantitative analysis of two independent binding modes of a small ligand responsible for DC-SIGN clustering.
Org Biomol Chem. (2016) 14: 335-44.

Pustylnikov S, Dave RS, Khan ZK, Porkolab V, Rashad AA, Hutchinson M, Fieschi F, Chaiken I, Jain P.
Short Communication: Inhibition of DC-SIGN-Mediated HIV-1 Infection by Complementary Actions of Dendritic Cell Receptor Antagonists and Env-Targeting Virus Inactivators.
AIDS Res Hum Retroviruses. (2016) 32: 93-100.

Giladi M, Almagor L, van Dijk L, Hiller R, Man P, Forest E and Khananshvili D.
Asymmetric preorganisation of inverted pair residues in the sodium-calcium exchanger.
Scientific Reports (2016) In press.

Muñoz-García JC, Chabrol E, Vivès RR, Thomas A, de Paz JL, Rojo J, Imberty A, Fieschi F, Nieto PM, Angulo J.
Langerin-heparin interaction: two binding sites for small and large ligands as revealed by a combination of NMR spectroscopy and cross-linking mapping experiments.
J Am Chem Soc. (2015) 137: 4100-10.

Chabrol E, Thépaut M, Dezutter-Dambuyant C, Vivès C, Marcoux J, Kahn R, Valladeau-Guilemond J, Vachette P, Durand D, Fieschi F.
Alteration of the langerin oligomerization state affects Birbeck granule formation.
Biophys J. (2015) 108: 666-77.

Ordanini S, Varga N, Porkolab V, Thépaut M, Belvisi L, Bertaglia A, Palmioli A, Berzi A, Trabattoni D, Clerici M, Fieschi F, Bernardi A.
Designing nanomolar antagonists of DC-SIGN-mediated HIV infection: ligand presentation using molecular rods.
Chem Commun (Camb). (2015) 51: 3816-9.

Lennartz F, Bengtsson A, Olsen RW, Joergensen L, Brown A, Remy L, Man P, Forest E, Barfod LK, Adams Y, Higgins MK, Jensen AT.
Mapping the Binding Site of a Cross-Reactive Plasmodium falciparum PfEMP1 Monoclonal Antibody Inhibitory of ICAM-1 Binding.
J Immunol. (2015) 195: 3273-83.

Beaumel S, Grunwald D, Fieschi F, Stasia MJ.
Identification of NOX2 regions for normal biosynthesis of cytochrome b558 in phagocytes highlighting essential residues for p22phox binding.
Biochem J. (2014) 464: 425-37.

Sutkeviciute I, Thépaut M, Sattin S, Berzi A, McGeagh J, Grudinin S, Weiser J, Le Roy A, Reina JJ, Rojo J, Clerici M, Bernardi A, Ebel C, Fieschi F.
Unique DC-SIGN clustering activity of a small glycomimetic: A lesson for ligand design.
ACS Chem Biol. (2014) 9: 1377-85.

Tomašić T, Hajšek D, Švajger U, Luzar J, Obermajer N, Petit-Haertlein I, Fieschi F, Anderluh M.
Monovalent mannose-based DC-SIGN antagonists: targeting the hydrophobic groove of the receptor.
Eur J Med Chem. (2014) 75: 308-26.

Varga N, Sutkeviciute I, Ribeiro-Viana R, Berzi A, Ramdasi R, Daghetti A, Vettoretti G, Amara A, Clerici M, Rojo J, Fieschi F, Bernardi A.
A multivalent inhibitor of the DC-SIGN dependent uptake of HIV-1 and Dengue virus.
Biomaterials (2014) 35: 4175-84.

Picciocchi A, Siaučiūnaiteė-Gaubard L, Petit-Hartlein I, Sadir R, Revilloud J, Caro L, Vivaudou M, Fieschi F, Moreau C, Vivès C.
C-terminal engineering of CXCL12 and CCL5 chemokines: functional characterization by electrophysiological recordings.
PLoS One (2014) 9: e87394.

Diotti RA, Mancini N, Clementi N, Sautto G, Moreno GJ, Criscuolo E, Cappelletti F, Man P, Forest E, Remy L, Giannecchini S, Clementi M, Burioni R.
Cloning of the first human anti-JCPyV/VP1 neutralizing monoclonal antibody: epitope definition and implications in risk stratification of patients under natalizumab therapy.
Antiviral Res. (2014) 108: 94-103.

Richichi B, Imberty A, Gillon E, Bosco R, Sutkeviciute I, Fieschi F, Nativi C.
Synthesis of a selective inhibitor of a fucose binding bacterial lectin from Burkholderia ambifaria.
Org Biomol Chem. (2013) 11: 4086-94.

Varga N, Sutkeviciute I, Guzzi C, McGeagh J, Petit-Haertlein I, Gugliotta S, Weiser J, Angulo J, Fieschi F, Bernardi A.
Selective targeting of dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) with mannose-based glycomimetics: synthesis and interaction studies of bis(benzylamide) derivatives of a pseudomannobioside.
Chemistry (2013) 19: 4786-97.

Thépaut M, Guzzi C, Sutkeviciute I, Sattin S, Ribeiro-Viana R, Varga N, Chabrol E, Rojo J, Bernardi A, Angulo J, Nieto PM, Fieschi F.
Structure of a glycomimetic ligand in the carbohydrate recognition domain of C-type lectin DC-SIGN. Structural requirements for selectivity and ligand design.
J Am Chem Soc. (2013) 135: 2518-29.

Bernardi A, Jiménez-Barbero J, Casnati A, De Castro C, Darbre T, Fieschi F, Finne J, Funken H, Jaeger KE, Lahmann M, Lindhorst TK, Marradi M, Messner P, Molinaro A, Murphy PV, Nativi C, Oscarson S, Penadés S, Peri F, Pieters RJ, Renaudet O, Reymond JL, Richichi B, Rojo J, Sansone F, Schäffer C, Turnbull WB, Velasco-Torrijos T, Vidal S, Vincent S, Wennekes T, Zuilhof H, Imberty A.
Multivalent glycoconjugates as anti-pathogenic agents.
Chem Soc Rev. (2013) 42: 4709-27.

Jacky BP, Garay PE, Dupuy J, Nelson JB, Cai B, Molina Y, Wang J, Steward LE, Broide RS, Francis J, Aoki KR, Stevens RC, Fernández-Salas E.
Identification of fibroblast growth factor receptor 3 (FGFR3) as a protein receptor for botulinum neurotoxin serotype A (BoNT/A).
PLoS Pathog. (2013) 9: e1003369.