Laboratoire Léon Brillouin

UMR12 CEA-CNRS, Bât. 563 CEA Saclay

91191 Gif sur Yvette Cedex, France

+33-169085241 llb-sec@cea.fr

BD diffusons les neutrons

Faits marquants scientifiques 2019

22 septembre 2019

Daniela Russo, Maya Dimova Lambreva, Christiane Alba Simionesco, Pierre Sebban,and Giuseppina Rea

Studies on the dynamical properties of photosynthetic membranes of land plants and purple bacteria have been previously performed by neutron spectroscopy, revealing a tight coupling between specific photochemical reactions and macromolecular dynamics. Here, we probed the intrinsic dynamics of biotechnologically useful mutants of the green alga Chlamydomonas reinhardtii by incoherent neutron scattering coupled with prompt chlorophyll fluorescence experiments. We brought to light that single amino acid replacements in the plastoquinone (PQ)-binding niche of the photosystem II D1 protein impair electron transport (ET) efficiency between quinones and confer increased flexibility to the host membranes, expanding to the entire cells. Hence, a more flexible environment in the PQ-binding niche has been associated to a less efficient ET.Asimilar function/dynamics relationship was also demonstrated in Rhodobacter sphaeroides reaction centers having inhibited ET, indicating that flexibility at the quinones region plays a crucial role in evolutionarily distant organisms. Instead, a different functional/dynamical correlation was observed in algal mutants hosting a single amino acid replacement residing in a D1 domain far from the PQ-binding niche. Noteworthy, this mutant displayed the highest degree of flexibility, and besides having a nativelike ET efficiency in physiological conditions, it acquired novel, to our knowledge, phenotypic traits enabling it to preserve a high maximal quantum yield of photosystem II photochemistry in extreme habitats. Overall, in the nanosecond timescale, the degree of the observed flexibility is related to the mutation site; in the picosecond timescale, we highlighted the presence of a more pronounced dynamic heterogeneity in all mutants compared to the native cells, which could be related to a marked chemically heterogeneous environment.

DOI : https://doi.org/10.1016/j.bpj.2019.03.029

 

27 août 2019

D. Russo, A. De Angelis, A. Paciaroni, B. Frick, N. de Sousa, F. R. Wurm, and J. Teixeira

We investigate the relaxation dynamics of proteinpolymer conjugates by neutron scattering spectroscopy to understand to which extent the coating of a protein by a polymer can replace water in promoting thermal structural fluctuations. For this purpose, we compare the dynamics of proteinpolymer mixtures to that of conjugates with a variable number of polymers covalently attached to the protein. Results show that the flexibility of the protein is larger in proteinpolymer mixtures than in native protein or in conjugates, even in the dry state. Upon hydration, both the native protein and the conjugate show equivalent dynamics, suggesting that the polymer grafted on the protein surface adsorbs all water molecules.

http://dx.doi.org/10.1021/acs.langmuir.8b03636

20 juin 2019

A. Theodoratou, L.-T. Lee, J. Oberdisse and A. Aubert-Pouëssel, Langmuir 35(20) (2019) 6620.

Abstract :

Nanofilms of about 2 nm thickness have been formed at the air–water interface using functionalized castor oil (ICO) with cross-linkable silylated groups. These hybrid films represent excellent candidates for replacing conventional polymeric materials in biomedical applications, but they need to be optimized in terms of biocompatibility, which is highly related to protein adsorption. Neutron reflectivity has been used to study the adsorption of two model proteins, bovine serum albumin and lysozyme, at the silylated oil (ICO)–water interface in the absence and presence of salt at physiologic ionic strength and pH and at different protein concentrations. These measurements are compared to adsorption at the air–water interface. While salt enhances adsorption by a similar degree at the air–water and oil–water interfaces, the impact of the oil film is significant with adsorption at the oil–water interface 3–4-fold higher compared to that at the air–water interface. Under these conditions, the concentration profiles of the adsorbed layers for both proteins indicate multilayer adsorption. The thickness of the outer layer (oil side) is close to the dimension of the minor axis of the protein molecule, ∼30 Å, suggesting a sideway orientation with the long axis parallel to the interface. The inner layer extends to 55–60 Å. Interestingly, in all cases, the composition of the oil film remains intact without significant protein penetration into the film. The optimal adsorption on these nanofilms, 1.7–2.0 mg·m-2, is comparable to the results obtained recently on thick solid cross-linked films using a quartz crystal microbalance and atomic force microscopy, showing in particular that adsorption at these ICO film interfaces under standard physiological conditions is nonspecific. These results furnish useful information toward the elaboration of vegetable oil-based nanofilms in direct nanoscale applications or as precursor films in the fabrication of thicker macroscopic films for biomedical applications.

https://doi.org/10.1021/acs.langmuir.9b00186

30 mai 2019

"Dynamics properties of photosynthetic microorganisms probed by incoherent neutron scattering"
Daniela Russo, Maya Dimova Lambreva, Christiane Alba Simionesco, Pierre Sebban, and Giuseppina Rea
Biophysical Journal, 116 (9) (2019) 1759-1768

Studies on the dynamical properties of photosynthetic membranes of land plants and purple bacteria have been previously performed by neutron spectroscopy, revealing a tight coupling between specific photochemical reactions and macromolecular dynamics. Here, we probed the intrinsic dynamics of biotechnologically useful mutants of the green alga Chlamydomonas reinhardtii by incoherent neutron scattering coupled with prompt chlorophyll fluorescence experiments. We brought to light that single amino acid replacements in the plastoquinone (PQ)-binding niche of the photosystem II D1 protein impair electron transport (ET) efficiency between quinones and confer increased flexibility to the host membranes, expanding to the entire cells. Hence, a more flexible environment in the PQ-binding niche has been associated to a less efficient ET.Asimilar function/dynamics relationship was also demonstrated in Rhodobacter sphaeroides reaction centers having inhibited ET, indicating that flexibility at the quinones region plays a crucial role in evolutionarily distant organisms. Instead, a different functional/dynamical correlation was observed in algal mutants hosting a single amino acid replacement residing in a D1 domain far from the PQ-binding niche. Noteworthy, this mutant displayed the highest degree of flexibility, and besides having a nativelike ET efficiency in physiological conditions, it acquired novel, to our knowledge, phenotypic traits enabling it to preserve a high maximal quantum yield of photosystem II photochemistry in extreme habitats. Overall, in the nanosecond timescale, the degree of the observed flexibility is related to the mutation site; in the picosecond timescale, we highlighted the presence of a more pronounced dynamic heterogeneity in all mutants compared to the native cells, which could be related to a marked chemically heterogeneous environment.

01 avril 2019

Andrés Marcoleta, Frank Wien, Véronique Arluison, Rosalba Lagos, Rafael Giraldo
Bacterial Amyloids (2019)

Amyloids are supramolecular protein assemblies based on fibrillar arrangements of β‐sheets that were first found as linked to neurodegenerative and systemic human diseases. However, there is now overwhelming evidence on alternative roles of amyloids as functional assemblies and as epigenetic determinants of beneficial traits, both in Fungi and Metazoa. Bacteria also use amyloids as functional devices, mainly as extracellular scaffolds in biofilms, but there is increasing evidence for functional roles of amyloids in the bacterial cytosol, and these have enabled to engineer minimal models of a ‘generic’ amyloid disease. Amyloids are thus key players in the physiology of bacteria and versatile building blocks in synthetic biology.

 
03 janvier 2019

Sophie Combet, Fabrice Cousin, Human Rezaei, Sylvie Noinville

Soluble oligomers of prion proteins (PrP), produced during amyloid aggregation, have emerged as the primary neurotoxic species, instead of the fibrillar end-products, in transmissible spongiform encephalopathies. However, whether the membrane is among their direct targets, that mediate the downstream adverse effects, remains a question of debate. Recently, questions arise from the formation of membrane-active oligomeric species generated during the β-aggregation pathway, either in solution, or in lipid environment. In the present study, we characterized membrane interaction of off-pathway oligomers from recombinant prion protein generated along the amyloid aggregation and compared to lipid-induced intermediates produced during lipid-accelerated fibrillation. Using calcein-leakage assay, we show that the soluble prion oligomers are the most potent in producing leakage with negatively charged vesicles. Binding affinities, conformational states, mode of action of the different PrP assemblies were determined by thioflavin T binding-static light scattering experiments on DOPC/DOPS vesicles, as well as by FTIR-ATR spectroscopy and specular neutron reflectivity onto the corresponding supported lipid bilayers. Our results indicate that the off-pathway PrP oligomers interact with lipid membrane via a distinct mechanism, compared to the inserted lipid-induced intermediates. Thus, separate neurotoxic mechanisms could exist following the puzzling intermediates generated in the different cell compartments. These results not only reveal an important regulation of lipid membrane on PrP behavior but may also provide clues for designing stage-specific and prion-targeted therapy.

https://doi.org/10.1016/j.bbamem.2018.12.001

 

20 juin 2019

A. Theodoratou, L.-T. Lee, J. Oberdisse and A. Aubert-Pouëssel, Langmuir 35(20) (2019) 6620.

Abstract :

Nanofilms of about 2 nm thickness have been formed at the air–water interface using functionalized castor oil (ICO) with cross-linkable silylated groups. These hybrid films represent excellent candidates for replacing conventional polymeric materials in biomedical applications, but they need to be optimized in terms of biocompatibility, which is highly related to protein adsorption. Neutron reflectivity has been used to study the adsorption of two model proteins, bovine serum albumin and lysozyme, at the silylated oil (ICO)–water interface in the absence and presence of salt at physiologic ionic strength and pH and at different protein concentrations. These measurements are compared to adsorption at the air–water interface. While salt enhances adsorption by a similar degree at the air–water and oil–water interfaces, the impact of the oil film is significant with adsorption at the oil–water interface 3–4-fold higher compared to that at the air–water interface. Under these conditions, the concentration profiles of the adsorbed layers for both proteins indicate multilayer adsorption. The thickness of the outer layer (oil side) is close to the dimension of the minor axis of the protein molecule, ∼30 Å, suggesting a sideway orientation with the long axis parallel to the interface. The inner layer extends to 55–60 Å. Interestingly, in all cases, the composition of the oil film remains intact without significant protein penetration into the film. The optimal adsorption on these nanofilms, 1.7–2.0 mg·m-2, is comparable to the results obtained recently on thick solid cross-linked films using a quartz crystal microbalance and atomic force microscopy, showing in particular that adsorption at these ICO film interfaces under standard physiological conditions is nonspecific. These results furnish useful information toward the elaboration of vegetable oil-based nanofilms in direct nanoscale applications or as precursor films in the fabrication of thicker macroscopic films for biomedical applications.

https://doi.org/10.1021/acs.langmuir.9b00186

30 mai 2019

"Dynamics properties of photosynthetic microorganisms probed by incoherent neutron scattering"
Daniela Russo, Maya Dimova Lambreva, Christiane Alba Simionesco, Pierre Sebban, and Giuseppina Rea
Biophysical Journal, 116 (9) (2019) 1759-1768

Studies on the dynamical properties of photosynthetic membranes of land plants and purple bacteria have been previously performed by neutron spectroscopy, revealing a tight coupling between specific photochemical reactions and macromolecular dynamics. Here, we probed the intrinsic dynamics of biotechnologically useful mutants of the green alga Chlamydomonas reinhardtii by incoherent neutron scattering coupled with prompt chlorophyll fluorescence experiments. We brought to light that single amino acid replacements in the plastoquinone (PQ)-binding niche of the photosystem II D1 protein impair electron transport (ET) efficiency between quinones and confer increased flexibility to the host membranes, expanding to the entire cells. Hence, a more flexible environment in the PQ-binding niche has been associated to a less efficient ET.Asimilar function/dynamics relationship was also demonstrated in Rhodobacter sphaeroides reaction centers having inhibited ET, indicating that flexibility at the quinones region plays a crucial role in evolutionarily distant organisms. Instead, a different functional/dynamical correlation was observed in algal mutants hosting a single amino acid replacement residing in a D1 domain far from the PQ-binding niche. Noteworthy, this mutant displayed the highest degree of flexibility, and besides having a nativelike ET efficiency in physiological conditions, it acquired novel, to our knowledge, phenotypic traits enabling it to preserve a high maximal quantum yield of photosystem II photochemistry in extreme habitats. Overall, in the nanosecond timescale, the degree of the observed flexibility is related to the mutation site; in the picosecond timescale, we highlighted the presence of a more pronounced dynamic heterogeneity in all mutants compared to the native cells, which could be related to a marked chemically heterogeneous environment.

21 avril 2019

Les membranes d’oxyde d’aluminium nanoporeuses sont des systèmes modèles permettant d’étudier le comportement de la matière sous confinement. Leurs utilisations pour des études de nano-moulage, de microfluidique ou en biologie sont ainsi nombreuses. Une équipe du laboratoire PHENIX (Université Pierre et Marie Curie) et du laboratoire Léon Brillouin (IRAMIS, CEA Saclay) ont étudié in-situ l’adsorption de polymères chargés dans ces matériaux modèles par une approche combinant la réflectivité de neutrons et la microscopie électronique.

04 mars 2019

Marguerite Léang, Didier Lairez, Fabrice Cousin, Frédérique Giorgiutti-Dauphiné, Ludovic Pauchard, Lay-Theng Lee

During evaporative drying of a colloidal dispersion, the structural behavior at the air-dispersion interface is of particular relevance to the understanding of the consolidation mechanism and the final structural and mechanical properties of the porous media. The drying-interface constitutes the region of initial drying-stress that, when accumulated over a critical thickness, leads to crack formation. This work presents an experimental study of top-down drying of colloidal silica dispersions with three different sizes (radius 5, 8 and 13 nm). Using specular neutron reflectivity, we focus on the structural evolution at the free drying-front of the dispersion with a macroscopic drying surface, and demonstrate the existence of a thick concentrated surface layer induced by heterogeneous evaporation. The reflectivity profile contains a strong structure peak due to scattering from particles in the interfacial region, from which the interparticle distance is deduced. A notable advantage of these measurements is the direct extraction of the corresponding dispersion concentration from the critical total reflection edge, providing a straightforward access to a structure-concentration relation during the drying process. The bulk reservoir of this experimental configuration renders it possible to verify the evaporation-diffusion balance to construct the surface layer and also to check reversibility of particle ordering. We follow the structural evolution of this surface layer from a sol to a soft wet-gel that is the precursor of a fragile skin, and the onset of significant particle aggregation that precedes formation of the wet-crust. Separate complementary measurements on the structural evolution in the bulk dispersion are also carried out by small angle neutron scattering, where the particle concentration is also extracted directly from the experimental curves. The two sets of data reveal similar structural evolution with concentration at the interface and in the bulk, and an increase in the degree of ordering with particle size.

https://pubs.acs.org/doi/pdfplus/10.1021/acs.langmuir.8b03772

25 septembre 2019

La physique de la matière condensée repose sur quelques concepts fondateurs, comme le paradigme de Néel (avec sa description des ordres magnétiques classiques), la théorie des liquides de Fermi (avec le concept de quasi-particule), et la théorie de Landau des transitions de phases (mettant en exergue le concept de brisure spontanée de symétrie). Toutefois, de nouvelles découvertes bouleversent ces dogmes, mettant en lumière le rôle des fortes corrélations entre quasi-particules, la découverte de transitions de phase topologiques, ou l’observation de nouveaux états de la matière.

Ce sont par exemple les liquides ou glaces de spin, les états de boucle de courant dans certains oxydes, ou la description de nouvelles transitions de phase, sans symétrie brisée, mettant en jeu le confinement/déconfinement d’objets topologiques comme les vortex. Ceci suscite et motive toujours plus avant l’étude de la matière et des "matériaux quantiques", pour acquérir de nouvelles connaissances fondamentales et en tirer le meilleur parti pour des applications diverses. Ces nouveaux états de la matière se rencontrent dans des conditions ultimes et notamment aux très basses températures, dans la gamme du milliKelvin (mK).

Dans cette optique, et pour accompagner cette recherche, notre équipe du LLB responsable de l’opération du diffractomètre de neutrons G4-1 (LLB-Orphée) a fait l’acquisition d’un nouvel appareil permettant de réaliser des mesures de diffraction des neutrons jusqu’à des températures ultra-basses (40 mK).

 

 

04 mai 2019

B. Annighöfer , A. Hélary, A. Brulet, A. Colas de la Noue, C. Loupiac and S. Combet
Review of Scientific Instruments 90 (2019) 025106

Abstract :

We report on a high pressure (HP) cell designed for the determination of the structure of molecular solutions by small-angle neutron scattering (SANS). The HP cell is fitted up with two thick metallic windows that make the device very resistant under hydrostatic pressures up to 600 MPa (or 6 kbar). The metallic windows are removable, offering the possibility to adapt the HP cell to a given study with the pressure desired on an appropriate spatial range to study the structure of various molecular solutions by SANS. In this context, we report the absorption, transmission, and scattering properties of different metallic windows. Finally, we describe, as a proof of principle, the solution structure changes of myoglobin, a small globular protein.

https://doi.org/10.1063/1.5051765

18 septembre 2019
Des chercheurs de l’I2BC@Saclay et de l’UMR NIMBE, en collaboration avec le Laboratoire Léon Brillouin (LLB), ont analysé la structure de la couronne composée de deux protéines modèles adsorbées sur des nanoparticules de silice, en utilisant la technique de diffusion des neutrons aux petits angles. Ils montrent que les structures formées sont de véritables nanoassemblages contrôlés, dans lesquels les protéines conservent leur forme.

 

27 août 2019

D. Russo, A. De Angelis, A. Paciaroni, B. Frick, N. de Sousa, F. R. Wurm, and J. Teixeira

We investigate the relaxation dynamics of proteinpolymer conjugates by neutron scattering spectroscopy to understand to which extent the coating of a protein by a polymer can replace water in promoting thermal structural fluctuations. For this purpose, we compare the dynamics of proteinpolymer mixtures to that of conjugates with a variable number of polymers covalently attached to the protein. Results show that the flexibility of the protein is larger in proteinpolymer mixtures than in native protein or in conjugates, even in the dry state. Upon hydration, both the native protein and the conjugate show equivalent dynamics, suggesting that the polymer grafted on the protein surface adsorbs all water molecules.

http://dx.doi.org/10.1021/acs.langmuir.8b03636

20 juin 2019

A. Theodoratou, L.-T. Lee, J. Oberdisse and A. Aubert-Pouëssel, Langmuir 35(20) (2019) 6620.

Abstract :

Nanofilms of about 2 nm thickness have been formed at the air–water interface using functionalized castor oil (ICO) with cross-linkable silylated groups. These hybrid films represent excellent candidates for replacing conventional polymeric materials in biomedical applications, but they need to be optimized in terms of biocompatibility, which is highly related to protein adsorption. Neutron reflectivity has been used to study the adsorption of two model proteins, bovine serum albumin and lysozyme, at the silylated oil (ICO)–water interface in the absence and presence of salt at physiologic ionic strength and pH and at different protein concentrations. These measurements are compared to adsorption at the air–water interface. While salt enhances adsorption by a similar degree at the air–water and oil–water interfaces, the impact of the oil film is significant with adsorption at the oil–water interface 3–4-fold higher compared to that at the air–water interface. Under these conditions, the concentration profiles of the adsorbed layers for both proteins indicate multilayer adsorption. The thickness of the outer layer (oil side) is close to the dimension of the minor axis of the protein molecule, ∼30 Å, suggesting a sideway orientation with the long axis parallel to the interface. The inner layer extends to 55–60 Å. Interestingly, in all cases, the composition of the oil film remains intact without significant protein penetration into the film. The optimal adsorption on these nanofilms, 1.7–2.0 mg·m-2, is comparable to the results obtained recently on thick solid cross-linked films using a quartz crystal microbalance and atomic force microscopy, showing in particular that adsorption at these ICO film interfaces under standard physiological conditions is nonspecific. These results furnish useful information toward the elaboration of vegetable oil-based nanofilms in direct nanoscale applications or as precursor films in the fabrication of thicker macroscopic films for biomedical applications.

https://doi.org/10.1021/acs.langmuir.9b00186

04 mars 2019

Marguerite Léang, Didier Lairez, Fabrice Cousin, Frédérique Giorgiutti-Dauphiné, Ludovic Pauchard, Lay-Theng Lee

During evaporative drying of a colloidal dispersion, the structural behavior at the air-dispersion interface is of particular relevance to the understanding of the consolidation mechanism and the final structural and mechanical properties of the porous media. The drying-interface constitutes the region of initial drying-stress that, when accumulated over a critical thickness, leads to crack formation. This work presents an experimental study of top-down drying of colloidal silica dispersions with three different sizes (radius 5, 8 and 13 nm). Using specular neutron reflectivity, we focus on the structural evolution at the free drying-front of the dispersion with a macroscopic drying surface, and demonstrate the existence of a thick concentrated surface layer induced by heterogeneous evaporation. The reflectivity profile contains a strong structure peak due to scattering from particles in the interfacial region, from which the interparticle distance is deduced. A notable advantage of these measurements is the direct extraction of the corresponding dispersion concentration from the critical total reflection edge, providing a straightforward access to a structure-concentration relation during the drying process. The bulk reservoir of this experimental configuration renders it possible to verify the evaporation-diffusion balance to construct the surface layer and also to check reversibility of particle ordering. We follow the structural evolution of this surface layer from a sol to a soft wet-gel that is the precursor of a fragile skin, and the onset of significant particle aggregation that precedes formation of the wet-crust. Separate complementary measurements on the structural evolution in the bulk dispersion are also carried out by small angle neutron scattering, where the particle concentration is also extracted directly from the experimental curves. The two sets of data reveal similar structural evolution with concentration at the interface and in the bulk, and an increase in the degree of ordering with particle size.

https://pubs.acs.org/doi/pdfplus/10.1021/acs.langmuir.8b03772

21 décembre 2019

M. Ruminy, S. Guitteny, J. Robert, L.-P. Regnault, M. Boehm, P. Steffens, H. Mutka, J. Ollivier, U. Stuhr, J. S. White, B. Roessli, L. Bovo, C. Decorse, M. K. Haas, R. J. Cava, I. Mirebeau, M. Kenzelmann, S. Petit, and T. Fennell

Tb2Ti2O7 presents an ongoing conundrum in the study of rare-earth pyrochlores. Despite the expectation that it should be the prototypical unfrustrated noncollinear Ising antiferromagnet on the pyrochlore lattice, it presents a puzzling correlated state that persists to the lowest temperatures. Effects which can reintroduce frustration or fluctuations are therefore sought, and quadrupolar operators have been implicated. One consequence of strong quadrupolar effects is the possible coupling of magnetic and lattice degrees of freedom, and it has previously been shown that a hybrid magnetoelastic mode with both magnetic and phononic character is formed in Tb2Ti2O7 by the interaction of a crystal field excitation with a transverse-acoustic phonon. Here, using polarized and unpolarized inelastic neutron scattering, we present a detailed characterization of the magnetic and phononic branches of this magnetoelastic mode, particularly with respect to their composition, the anisotropy of any magnetic fluctuations, and also the temperature dependence of the different types of fluctuation that are involved. We also examine the dispersion relations of the exciton branches that develop from the crystal field excitation in the same temperature regime that the coupled mode appears, and find three quasidispersionless branches where four are expected, each with a distinctive structure factor indicating that they are nonetheless cooperative excitations. We interpret the overall structure of the spectrum as containing four branches, one hybridized with the phonons and gaining a strong dispersion, and three remaining dispersionless.

https ://doi.org/10.1103/PhysRevB.99.224431

18 septembre 2019
Des chercheurs de l’I2BC@Saclay et de l’UMR NIMBE, en collaboration avec le Laboratoire Léon Brillouin (LLB), ont analysé la structure de la couronne composée de deux protéines modèles adsorbées sur des nanoparticules de silice, en utilisant la technique de diffusion des neutrons aux petits angles. Ils montrent que les structures formées sont de véritables nanoassemblages contrôlés, dans lesquels les protéines conservent leur forme.

 

22 septembre 2019

Daniela Russo, Maya Dimova Lambreva, Christiane Alba Simionesco, Pierre Sebban,and Giuseppina Rea

Studies on the dynamical properties of photosynthetic membranes of land plants and purple bacteria have been previously performed by neutron spectroscopy, revealing a tight coupling between specific photochemical reactions and macromolecular dynamics. Here, we probed the intrinsic dynamics of biotechnologically useful mutants of the green alga Chlamydomonas reinhardtii by incoherent neutron scattering coupled with prompt chlorophyll fluorescence experiments. We brought to light that single amino acid replacements in the plastoquinone (PQ)-binding niche of the photosystem II D1 protein impair electron transport (ET) efficiency between quinones and confer increased flexibility to the host membranes, expanding to the entire cells. Hence, a more flexible environment in the PQ-binding niche has been associated to a less efficient ET.Asimilar function/dynamics relationship was also demonstrated in Rhodobacter sphaeroides reaction centers having inhibited ET, indicating that flexibility at the quinones region plays a crucial role in evolutionarily distant organisms. Instead, a different functional/dynamical correlation was observed in algal mutants hosting a single amino acid replacement residing in a D1 domain far from the PQ-binding niche. Noteworthy, this mutant displayed the highest degree of flexibility, and besides having a nativelike ET efficiency in physiological conditions, it acquired novel, to our knowledge, phenotypic traits enabling it to preserve a high maximal quantum yield of photosystem II photochemistry in extreme habitats. Overall, in the nanosecond timescale, the degree of the observed flexibility is related to the mutation site; in the picosecond timescale, we highlighted the presence of a more pronounced dynamic heterogeneity in all mutants compared to the native cells, which could be related to a marked chemically heterogeneous environment.

DOI : https://doi.org/10.1016/j.bpj.2019.03.029

 

27 août 2019

D. Russo, A. De Angelis, A. Paciaroni, B. Frick, N. de Sousa, F. R. Wurm, and J. Teixeira

We investigate the relaxation dynamics of proteinpolymer conjugates by neutron scattering spectroscopy to understand to which extent the coating of a protein by a polymer can replace water in promoting thermal structural fluctuations. For this purpose, we compare the dynamics of proteinpolymer mixtures to that of conjugates with a variable number of polymers covalently attached to the protein. Results show that the flexibility of the protein is larger in proteinpolymer mixtures than in native protein or in conjugates, even in the dry state. Upon hydration, both the native protein and the conjugate show equivalent dynamics, suggesting that the polymer grafted on the protein surface adsorbs all water molecules.

http://dx.doi.org/10.1021/acs.langmuir.8b03636

20 juin 2019

A. Theodoratou, L.-T. Lee, J. Oberdisse and A. Aubert-Pouëssel, Langmuir 35(20) (2019) 6620.

Abstract :

Nanofilms of about 2 nm thickness have been formed at the air–water interface using functionalized castor oil (ICO) with cross-linkable silylated groups. These hybrid films represent excellent candidates for replacing conventional polymeric materials in biomedical applications, but they need to be optimized in terms of biocompatibility, which is highly related to protein adsorption. Neutron reflectivity has been used to study the adsorption of two model proteins, bovine serum albumin and lysozyme, at the silylated oil (ICO)–water interface in the absence and presence of salt at physiologic ionic strength and pH and at different protein concentrations. These measurements are compared to adsorption at the air–water interface. While salt enhances adsorption by a similar degree at the air–water and oil–water interfaces, the impact of the oil film is significant with adsorption at the oil–water interface 3–4-fold higher compared to that at the air–water interface. Under these conditions, the concentration profiles of the adsorbed layers for both proteins indicate multilayer adsorption. The thickness of the outer layer (oil side) is close to the dimension of the minor axis of the protein molecule, ∼30 Å, suggesting a sideway orientation with the long axis parallel to the interface. The inner layer extends to 55–60 Å. Interestingly, in all cases, the composition of the oil film remains intact without significant protein penetration into the film. The optimal adsorption on these nanofilms, 1.7–2.0 mg·m-2, is comparable to the results obtained recently on thick solid cross-linked films using a quartz crystal microbalance and atomic force microscopy, showing in particular that adsorption at these ICO film interfaces under standard physiological conditions is nonspecific. These results furnish useful information toward the elaboration of vegetable oil-based nanofilms in direct nanoscale applications or as precursor films in the fabrication of thicker macroscopic films for biomedical applications.

https://doi.org/10.1021/acs.langmuir.9b00186

30 mai 2019

"Dynamics properties of photosynthetic microorganisms probed by incoherent neutron scattering"
Daniela Russo, Maya Dimova Lambreva, Christiane Alba Simionesco, Pierre Sebban, and Giuseppina Rea
Biophysical Journal, 116 (9) (2019) 1759-1768

Studies on the dynamical properties of photosynthetic membranes of land plants and purple bacteria have been previously performed by neutron spectroscopy, revealing a tight coupling between specific photochemical reactions and macromolecular dynamics. Here, we probed the intrinsic dynamics of biotechnologically useful mutants of the green alga Chlamydomonas reinhardtii by incoherent neutron scattering coupled with prompt chlorophyll fluorescence experiments. We brought to light that single amino acid replacements in the plastoquinone (PQ)-binding niche of the photosystem II D1 protein impair electron transport (ET) efficiency between quinones and confer increased flexibility to the host membranes, expanding to the entire cells. Hence, a more flexible environment in the PQ-binding niche has been associated to a less efficient ET.Asimilar function/dynamics relationship was also demonstrated in Rhodobacter sphaeroides reaction centers having inhibited ET, indicating that flexibility at the quinones region plays a crucial role in evolutionarily distant organisms. Instead, a different functional/dynamical correlation was observed in algal mutants hosting a single amino acid replacement residing in a D1 domain far from the PQ-binding niche. Noteworthy, this mutant displayed the highest degree of flexibility, and besides having a nativelike ET efficiency in physiological conditions, it acquired novel, to our knowledge, phenotypic traits enabling it to preserve a high maximal quantum yield of photosystem II photochemistry in extreme habitats. Overall, in the nanosecond timescale, the degree of the observed flexibility is related to the mutation site; in the picosecond timescale, we highlighted the presence of a more pronounced dynamic heterogeneity in all mutants compared to the native cells, which could be related to a marked chemically heterogeneous environment.

04 mai 2019

B. Annighöfer , A. Hélary, A. Brulet, A. Colas de la Noue, C. Loupiac and S. Combet
Review of Scientific Instruments 90 (2019) 025106

Abstract :

We report on a high pressure (HP) cell designed for the determination of the structure of molecular solutions by small-angle neutron scattering (SANS). The HP cell is fitted up with two thick metallic windows that make the device very resistant under hydrostatic pressures up to 600 MPa (or 6 kbar). The metallic windows are removable, offering the possibility to adapt the HP cell to a given study with the pressure desired on an appropriate spatial range to study the structure of various molecular solutions by SANS. In this context, we report the absorption, transmission, and scattering properties of different metallic windows. Finally, we describe, as a proof of principle, the solution structure changes of myoglobin, a small globular protein.

https://doi.org/10.1063/1.5051765

01 avril 2019

Andrés Marcoleta, Frank Wien, Véronique Arluison, Rosalba Lagos, Rafael Giraldo
Bacterial Amyloids (2019)

Amyloids are supramolecular protein assemblies based on fibrillar arrangements of β‐sheets that were first found as linked to neurodegenerative and systemic human diseases. However, there is now overwhelming evidence on alternative roles of amyloids as functional assemblies and as epigenetic determinants of beneficial traits, both in Fungi and Metazoa. Bacteria also use amyloids as functional devices, mainly as extracellular scaffolds in biofilms, but there is increasing evidence for functional roles of amyloids in the bacterial cytosol, and these have enabled to engineer minimal models of a ‘generic’ amyloid disease. Amyloids are thus key players in the physiology of bacteria and versatile building blocks in synthetic biology.

 
15 février 2019

Burkhard Annighöfer, Arnaud Hélary, Annie Brûlet, Alexandre Colas de la Noue, Camille Loupiac, and Sophie Combet

We report on a high pressure (HP) cell designed for the determination of the structure of molecular solutions by small-angle neutron scattering (SANS). The HP cell is fitted up with two thick metallic windows that make the device very resistant under hydrostatic pressures up to 600 MPa (or 6 kbar). The metallic windows are removable, offering the possibility to adapt the HP cell to a given study with the pressure desired on an appropriate spatial range to study the structure of various molecular solutions by SANS. In this context, we report the absorption, transmission, and scattering properties of different metallic windows. Finally, we describe, as a proof of principle, the solution structure changes of myoglobin, a small globular protein.

https://doi.org/10.1063/1.5051765

03 janvier 2019

Sophie Combet, Fabrice Cousin, Human Rezaei, Sylvie Noinville

Soluble oligomers of prion proteins (PrP), produced during amyloid aggregation, have emerged as the primary neurotoxic species, instead of the fibrillar end-products, in transmissible spongiform encephalopathies. However, whether the membrane is among their direct targets, that mediate the downstream adverse effects, remains a question of debate. Recently, questions arise from the formation of membrane-active oligomeric species generated during the β-aggregation pathway, either in solution, or in lipid environment. In the present study, we characterized membrane interaction of off-pathway oligomers from recombinant prion protein generated along the amyloid aggregation and compared to lipid-induced intermediates produced during lipid-accelerated fibrillation. Using calcein-leakage assay, we show that the soluble prion oligomers are the most potent in producing leakage with negatively charged vesicles. Binding affinities, conformational states, mode of action of the different PrP assemblies were determined by thioflavin T binding-static light scattering experiments on DOPC/DOPS vesicles, as well as by FTIR-ATR spectroscopy and specular neutron reflectivity onto the corresponding supported lipid bilayers. Our results indicate that the off-pathway PrP oligomers interact with lipid membrane via a distinct mechanism, compared to the inserted lipid-induced intermediates. Thus, separate neurotoxic mechanisms could exist following the puzzling intermediates generated in the different cell compartments. These results not only reveal an important regulation of lipid membrane on PrP behavior but may also provide clues for designing stage-specific and prion-targeted therapy.

https://doi.org/10.1016/j.bbamem.2018.12.001

 

05 mars 2019

Annelies Sels, Giovanni Salassa, Fabrice Cousin, Lay-Theng Lee, Thomas Bürgi,
Nanoscale 26 (2018) 12754.

Aromatic dithiol linkers were used to prepare aggregates of Au25(SR)18 clusters (SR: thiolate) via ligand exchange reactions. Fractions of different aggregate sizes were separated by size exclusion chromatography (SEC). The aggregates were characterized by UV-vis absorption spectroscopy, matrix assisted laser desorption ionization (MALDI) mass spectrometry, nuclear magnetic resonance (NMR) spectroscopy (including diffusion-ordered spectroscopy, DOSY) and small angle X-ray scattering (SAXS).

At 2:1 cluster: dithiol ratio, small aggregates (dimers, trimers) and also larger aggregates consisting of 10-20 Au25 clusters were formed, according to DOSY, besides unreacted (monomeric) Au25(SR)18. MALDI mass spectrometry shows signals consistent with dimers and trimers (doubly charged). The SAXS curves for the small aggregates can be well fitted by a pearl-necklace model. For the bigger aggregates the SAXS curves evidence a characteristic separation distance between the clusters within the aggregates, which is imposed by the length of the linker. The SAXS curves of these larger aggregates can be well fitted with a core-shell sphere model with a sticky hard-sphere structure factor, in agreement with closely packed aggregates.

The absorption spectra of smaller aggregates resemble the one of individual Au25(SR)18 clusters; however, and most importantly, the larger aggregates show completely different, less structured spectra with a new band emerging at 840 nm. We assign this drastic change in the absorption spectra and the new band to the electronic coupling between the clusters through the all aromatic linker. In accordance with this view, aggregates formed with a linker containing methylene groups, thus breaking conjugation, do not show the band at 840 nm. By addition of monothiols to the larger aggregates their size can be reduced through an “unlinking” reaction. This reaction also affects the band at 840 nm, which moves to higher energy when reducing the aggregate size, as would be expected within a particle in a box model. The electronic coupling between the clusters through the linker is the basis for future applications in nanoelectronics.

http://dx.doi.org/10.1039/C8NR01902A

25 novembre 2019

A. S. Sukhanov, Praveen Vir, A. S. Cameron, H. C. Wu, N. Martin, S. Mühlbauer, A. Heinemann, H. D. Yang, C. Felser, and D. S. Inosov

The cubic chiral helimagnets with the P213 space group represent a group of compounds in which the stable skyrmion-lattice state is experimentally observed. The key parameter that controls the energy landscape of such systems and determines the emergence of a topologically nontrivial magnetic structure is the Dzyaloshinskii-Moriya interaction (DMI). Chemical substitution is recognized as a convenient instrument to tune the DMI in real materials and has been successfully utilized in studies of a number of chiral magnets, such as MnSi, FeGe, MnGe, and others. In our study, we applied small-angle neutron scattering to investigate how chemical substitution influences the skyrmionic properties of an insulating helimagnet Cu2OSeO3 when Cu ions are replaced by either Zn or Ni. Our results demonstrate that the DMI is enhanced in the Ni-substituted compounds (Cu,Ni)2OSeO3, but weakened in (Cu,Zn)2OSeO3. The observed changes in the DMI strength are reflected in the magnitude of the spin-spiral propagation vector and the temperature stability of the skyrmion phase.

https://doi.org/10.1103/PhysRevB.100.184408

23 octobre 2019

N. Martin, I. Mirebeau, C. Franz, G. Chaboussant, L. N. Fomicheva, and A. V. Tsvyashchenko

We study the helimagnetic ground state of the MnGe cubic alloy using small-angle neutron scattering and a high-resolution method, the so-called MIEZE spectroscopy. Upon cooling below the Néel temperature TN = 170(5) K, we observe the proliferation of long-wavelength gapless spin fluctuations, concomitant with a continuous evolution of the helical correlation length. These fluctuations disappear at Tcom = 32(5) K when the helical period becomes commensurate with the lattice. We propose to describe this intermediate phase as a soliton lattice, promoting nonlinear collective modes, or phasons, over a large temperature interval. We discuss the possible relevance of our results to the previously observed magnetotransport anomalies.

https://doi.org/10.1103/PhysRevB.99.100402

04 mars 2019

Marguerite Léang, Didier Lairez, Fabrice Cousin, Frédérique Giorgiutti-Dauphiné, Ludovic Pauchard, Lay-Theng Lee

During evaporative drying of a colloidal dispersion, the structural behavior at the air-dispersion interface is of particular relevance to the understanding of the consolidation mechanism and the final structural and mechanical properties of the porous media. The drying-interface constitutes the region of initial drying-stress that, when accumulated over a critical thickness, leads to crack formation. This work presents an experimental study of top-down drying of colloidal silica dispersions with three different sizes (radius 5, 8 and 13 nm). Using specular neutron reflectivity, we focus on the structural evolution at the free drying-front of the dispersion with a macroscopic drying surface, and demonstrate the existence of a thick concentrated surface layer induced by heterogeneous evaporation. The reflectivity profile contains a strong structure peak due to scattering from particles in the interfacial region, from which the interparticle distance is deduced. A notable advantage of these measurements is the direct extraction of the corresponding dispersion concentration from the critical total reflection edge, providing a straightforward access to a structure-concentration relation during the drying process. The bulk reservoir of this experimental configuration renders it possible to verify the evaporation-diffusion balance to construct the surface layer and also to check reversibility of particle ordering. We follow the structural evolution of this surface layer from a sol to a soft wet-gel that is the precursor of a fragile skin, and the onset of significant particle aggregation that precedes formation of the wet-crust. Separate complementary measurements on the structural evolution in the bulk dispersion are also carried out by small angle neutron scattering, where the particle concentration is also extracted directly from the experimental curves. The two sets of data reveal similar structural evolution with concentration at the interface and in the bulk, and an increase in the degree of ordering with particle size.

https://pubs.acs.org/doi/pdfplus/10.1021/acs.langmuir.8b03772

25 janvier 2019

M. Deutsch, W. Peng, P. Foury-Leylekian, V. Balédent, S. Chattopadhyay, M. T. Fernandez-Diaz, T. C. Hansen, A. Forget, D. Colson, M. Greenblatt, M.-B. Lepetit, S. Petit, and I. Mirebeau

The magnetic structure of TbMn2O5 and DyMn2O5 multiferroics has been studied by high-pressure neutron diffraction in a large pressure range up to 6.6 GPa. In both cases, we observe a pressure-induced commensurate magnetic phase with propagation vector ( 1/2 0 1/2 ), growing with pressure at the expense of the ambient pressure phases. Being previously observed in YMn2O5 and PrMn2O5, this phase is most likely a generic feature of the RMn2O5 multiferroic family. A simple model is proposed to explain qualitatively the emergence of this pressure-induced phase. Differences between TbMn2O5 and DyMn2O5 behaviors at ambient and low pressures provide clues on the interaction scheme.

DOI : http://dx.doi.org/10.1103/PhysRevB.98.024408

21 décembre 2019

M. Ruminy, S. Guitteny, J. Robert, L.-P. Regnault, M. Boehm, P. Steffens, H. Mutka, J. Ollivier, U. Stuhr, J. S. White, B. Roessli, L. Bovo, C. Decorse, M. K. Haas, R. J. Cava, I. Mirebeau, M. Kenzelmann, S. Petit, and T. Fennell

Tb2Ti2O7 presents an ongoing conundrum in the study of rare-earth pyrochlores. Despite the expectation that it should be the prototypical unfrustrated noncollinear Ising antiferromagnet on the pyrochlore lattice, it presents a puzzling correlated state that persists to the lowest temperatures. Effects which can reintroduce frustration or fluctuations are therefore sought, and quadrupolar operators have been implicated. One consequence of strong quadrupolar effects is the possible coupling of magnetic and lattice degrees of freedom, and it has previously been shown that a hybrid magnetoelastic mode with both magnetic and phononic character is formed in Tb2Ti2O7 by the interaction of a crystal field excitation with a transverse-acoustic phonon. Here, using polarized and unpolarized inelastic neutron scattering, we present a detailed characterization of the magnetic and phononic branches of this magnetoelastic mode, particularly with respect to their composition, the anisotropy of any magnetic fluctuations, and also the temperature dependence of the different types of fluctuation that are involved. We also examine the dispersion relations of the exciton branches that develop from the crystal field excitation in the same temperature regime that the coupled mode appears, and find three quasidispersionless branches where four are expected, each with a distinctive structure factor indicating that they are nonetheless cooperative excitations. We interpret the overall structure of the spectrum as containing four branches, one hybridized with the phonons and gaining a strong dispersion, and three remaining dispersionless.

https ://doi.org/10.1103/PhysRevB.99.224431


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