title: Single-chain protein mimetics of the N-terminal heptad-repeat region of gp41 with potential as anti-HIV-1 drugs.
abstract: During HIV-1 fusion to the host cell membrane, the N-terminal heptad repeat (NHR) and the C-terminal heptad repeat (CHR) of the envelope subunit gp41 become transiently exposed and accessible to fusion inhibitors or Abs. In this process, the NHR region adopts a trimeric coiled-coil conformation that can be a target for therapeutic intervention. Here, we present an approach to rationally design single-chain protein constructs that mimic the NHR coiled-coil surface. The proteins were built by connecting with short loops two parallel NHR helices and an antiparallel one with the inverse sequence followed by engineering of stabilizing interactions. The constructs were expressed in Escherichia coli, purified with high yield, and folded as highly stable helical coiled coils. The crystal structure of one of the constructs confirmed the predicted fold and its ability to accurately mimic an exposed gp41 NHR surface. These single-chain proteins bound to synthetic CHR peptides with very high affinity, and furthermore, they showed broad inhibitory activity of HIV-1 fusion on various pseudoviruses and primary isolates.
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain;
authors.forename: Sara
authors.initials: S
authors.lastname: Crespillo
authors.affiliation: Department of Chemistry and Physics, Agrifood Campus of International Excellence (ceiA3), University of Almería, 04120 Almería, Spain;
authors.forename: Ana
authors.initials: A
authors.lastname: Cámara-Artigas
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain;
authors.forename: Salvador
authors.initials: S
authors.lastname: Casares
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain;
authors.forename: Bertrand
authors.initials: B
authors.lastname: Morel
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain;
authors.forename: Eva S
authors.initials: ES
authors.lastname: Cobos
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain;
authors.forename: Pedro L
authors.initials: PL
authors.lastname: Mateo
authors.affiliation: PX'Therapeutics, 38040 Grenoble Cedex 9, France;
authors.forename: Nicolas
authors.initials: N
authors.lastname: Mouz
authors.affiliation: PX'Therapeutics, 38040 Grenoble Cedex 9, France;
authors.forename: Christophe E
authors.initials: CE
authors.lastname: Martin
authors.affiliation: PX'Therapeutics, 38040 Grenoble Cedex 9, France;
authors.forename: Marie G
authors.initials: MG
authors.lastname: Roger
authors.affiliation: Sanofi Pasteur S.A., Campus Mérieux, 69280 Marcy l'Etoile, France; and.
authors.forename: Raphaelle
authors.initials: R
authors.lastname: El Habib
authors.affiliation: INSERM U1109, Fédération de Médecine Translationnelle de Strasbourg, Université de Strasbourg, 67000 Strasbourg, France.
authors.forename: Bin
authors.initials: B
authors.lastname: Su
authors.affiliation: INSERM U1109, Fédération de Médecine Translationnelle de Strasbourg, Université de Strasbourg, 67000 Strasbourg, France.
authors.forename: Christiane
authors.initials: C
authors.lastname: Moog
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain; conejero@ugr.es.
authors.forename: Francisco
authors.initials: F
authors.lastname: Conejero-Lara
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D055592:Biophysical Phenomena; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D018716:Molecular Mimicry
chemical_list: D019380:Anti-HIV Agents; D015700:HIV Envelope Protein gp41
keywords: HIV-1 neutralization; X-ray crystallography; coiled coil; drug design; fusion inhibitor