title: Self-association process of a peptide in solution: from beta-sheet filaments to large embedded nanotubes.
abstract: Lanreotide is a synthetic octapeptide used in the therapy against acromegaly. When mixed with pure water at 10% (w/w), Lanreotide (acetate salt) forms liquid crystalline and monodisperse nanotubes with a radius of 120 A. The molecular and supramolecular organization of these structures has been determined in a previous work as relying on the lateral association of 26 beta-sheet filaments made of peptide noncovalent dimers, the basic building blocks. The work presented here has been devoted to the corresponding self-association mechanisms, through the characterization of the Lanreotide structures formed in water, as a function of peptide (acetate salt) concentration (from 2% to 70% (w/w)) and temperature (from 15 degrees C to 70 degrees C). The corresponding states of water were also identified and quantified from the thermal behavior of water in the Lanreotide mixtures. At room temperature and below 3% (w/w) Lanreotide acetate in water, soluble aggregates were detected. From 3% to 20% (w/w) long individual and monodisperse nanotubes crystallized in a hexagonal lattice were evidenced. Their molecular and supramolecular organizations are identical to the ones characterized for the 10% (w/w) sample. Heating induces the dissolution of the nanotubes into soluble aggregates of the same structural characteristics as the room temperature ones. The solubilization temperature increases from 20 degrees C to 70 degrees C with the peptide concentration and reaches a plateau between 15% and 25% (w/w) in peptide. These aggregates are proposed to be the beta-sheet filaments that self-associate to build the walls of the nanotubes. Above 20% (w/w) of Lanreotide acetate in water, polydisperse embedded nanotubes are formed and the hexagonal lattice is lost. These embedded nanotubes exhibit the same molecular and supramolecular organizations as the individual monodisperse nanotubes formed at lower peptide concentration. The embedded nanotubes do not melt in the range of temperature studied indicating a higher thermodynamic stability than individual nanotubes. In parallel, the thermal behaviors of water in mixtures containing 2-80% (w/w) in peptide have been studied by differential scanning calorimetry, and three different types of water were characterized: 1), bulk water melting at 0 degrees C, 2), nonfreezing water, and 3), interfacial water melting below 0 degrees C. The domains of existence and coexistence of these different water states are related to the different Lanreotide supramolecular structures. All these results were compiled into a binary Lanreotide-water phase diagram and allowed to propose a self-association mechanism of Lanreotide filaments into monodisperse individual nanotubes and embedded nanotubes.
authors.affiliation: Unité Mixte de Recherche Centre National de la Recherche Scientifique 8612, Faculté de Pharmacie, Châtenay-Malabry, France.
authors.forename: C
authors.initials: C
authors.lastname: Valéry
authors.forename: F
authors.initials: F
authors.lastname: Artzner
authors.forename: B
authors.initials: B
authors.lastname: Robert
authors.forename: T
authors.initials: T
authors.lastname: Gulick
authors.forename: G
authors.initials: G
authors.lastname: Keller
authors.forename: C
authors.initials: C
authors.lastname: Grabielle-Madelmont
authors.forename: M-L
authors.initials: ML
authors.lastname: Torres
authors.forename: R
authors.initials: R
authors.lastname: Cherif-Cheikh
authors.forename: M
authors.initials: M
authors.lastname: Paternostre
mesh_terms: D002152:Calorimetry, Differential Scanning; D008854:Microscopy, Electron; D043942:Nanotubes; D010455:Peptides / Q000737:chemistry*; D010456:Peptides, Cyclic / Q000737:chemistry*; D044367:Phase Transition; D012996:Solutions / Q000737:chemistry; D013004:Somatostatin / Q000031:analogs & derivatives / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D013816:Thermodynamics; D014961:X-Ray Diffraction
chemical_list: D010455:Peptides; D010456:Peptides, Cyclic; D012996:Solutions; C060347:lanreotide; D013004:Somatostatin