Modeling procyanidin self-association processes and understanding their micellar organization: a study by diffusion NMR and molecular mechanics

The colloidal behavior of eight synthetic procyanidins (three monomers, four dimers, and a trimer) has been investigated in water or in a winelike medium using DOSY NMR spectroscopy and molecular dynamics simulations. Different behavior was observed for monomers and oligomers. Monomers self-associate with a high affinity constant (37-53 M(-1)) to form micelles at low cmc (critical micelle concentration) values (1-5 g.L(-1)). These micelles undergo a time-dependent coalescence process to form hazes and precipitates. As for dimers and the trimer, self-association also occurs but with a lower affinity (approximately 6 M(-1)) and at higher cmc values (10-20 g.L(-1)) to form small micelles (<5 nm) that remain stable throughout the experiment. The presence of 10% ethanol does not significantly affect the self-association constant for monomers and oligomers but increases their cmc values by approximately 50% and decreases the micelle size by a factor 2. However, the presence of 20 mM NaCl appears to negate the effect of ethanol. This study helps to clarify the role of procyanidin monomers versus oligomers in wine turbidity and demonstrates that procyanidin oligomers are fully available to interact with saliva proteins.     

NMR methods for studying the structure and dynamics of oncogenic and antihistaminic peptides in biomembranes

We present several applications of both wide-line and magic angle spinning (MAS) solid-state NMR of bicelles in which are embedded fragments of a tyrosine kinase receptor or enkephalins. The magnetically orientable bicelle membranes are shown to be of particular interest for studying the functional properties of lipids and proteins in a state that is very close to their natural environment. Quadrupolar, dipolar and chemical shielding interactions can be used to determine minute alterations of internal membrane dynamics and the orientation of peptides with respect to the membrane plane. MAS of bicelles can in turn lead to high-resolution proton spectra of hydrated membranes. Using deuterium-proton contrast methods one can then obtain pseudo-high-resolution proton spectra of peptides or proteins embedded in deuterated membranes and determine their atomic 3D structure using quasi-conventional liquid-state NMR methods.     

Sequestration of Proteins by Fatty Acid Coacervates for Their Encapsulation within Vesicles

Encapsulating biological materials in lipid vesicles is of interest for mimicking cells; however, except in some particular cases, such processes do not occur spontaneously. Herein, we developed a simple and robust method for encapsulating proteins in fatty acid vesicles in high yields. Fatty acid based, membrane‐free coacervates spontaneously sequester proteins and can reversibly form membranous vesicles upon varying the pH value, the precrowding feature in coacervates allowing for protein encapsulation within vesicles. We then produced enzyme‐enriched vesicles and show that enzymatic reactions can occur in these micrometric capsules. This work could be of interest in the field of synthetic biology for building microreactors.     

Structural and conformational analysis of two native procyanidin trimers

The structure and conformation of two native procyanidin trimers in water have been determined using 2D NMR and molecular mechanics. The results show the existence of four rotameric forms, one of which is predominant (60 to 80%). These four rotamers are shown to be in slow to intermediate exchange on the NMR timescale. Both trimers, whose structures vary owing to a different substitution of one carbon atom, adopt conformations in which stacking between different phenolic rings is favored.     

Toward bicelle stability with ether-linked phospholipids: temperature, composition, and hydration diagrams by 2H and 31P solid-state NMR

Phosphorus and deuterium wide line NMR was used to determine diagrams of binary mixtures of 1,2-di-O-tetradecyl-sn-glycero-3-phosphocholine (DIOMPC) and 1,2-di-O-hexyl-sn-glycero-3-phosphocholine (DIOHPC) ether-phospholipids. By varying the hydration, h, the temperature, T, and the mole fraction, X, of long-chain ether-phospholipids, we delineated the conditions for which such systems are oriented by the magnetic field, in the presence of 100 mM KCl. The 3D domain is found for X = 62-90%, T = 27-50 degrees C, and h = 70-98%. At 80% hydration, the domain shape (X = 70-90% and T = 27-42 degrees C) is close to that already observed for ester-phospholipids mixtures (Raffard, G.; Steinbruckner, S.; Arnold, A.; Davis, J. H.; Dufourc, E. J. Langmuir 2000, 16, 7655-7662) where disc-shaped bicelles of 300-600 A have been found by electron microscopy (Arnold, A.; Labrot, T.; Oda, R.; Dufourc, E. J. Biophys. J. 2002, 83, 2667-2680). Systems made of ether-linked lipids are much more stable on time and acidic conditions than those made of ester lipids. Assuming that the disc-shaped species are also found with ether lipids, their diameter as determined from integration of phosphorus NMR lines ranges from 240 to 440 A +/- 10%; it is generally independent of hydration and temperature but decreases with decreasing long-chain lipid content, X. The structure and the dynamics of water in the DIOMPC-DIOHPC were characterized by (2)H NMR. Water exchanges between the membrane surface where it is bound and a bulk isotropic pool lead to an average ordered state for temperatures in the bicelle region and above, thus offering a larger thermal span for structural studies of dissolved molecules.     

NMR of human saliva protein/wine tannin complexes. Towards deciphering astringency with physico-chemical tools

In the present work, we used NMR (chemical shift variations and diffusion measurement) to determine the affinity and the number of sites of four different 4-8 procyanidin dimers for a model PRP peptide. A number of three binding sites has been identified for every tannin, but significant affinity differences are highlighted from one to another dimer (a factor 10 approximately) that could be related to their specific 3D-structure.     

Liquid crystalline polymers as stationary phases. IV. Chemical bonding and immobilization of the polymer on silica characterization by solid-state nuclear magnetic resonance spectroscopy

Chemical bonding reaction and immobilization through low energy radiation (heating) have been investigated to fix a side-chain liquid crystalline polymer (SC-LCP) on silica particles in order to use the resulting modified silica in normal-phase HPLC. Highly stable chromatographic stationary phases are observed under excellent polymer solvent flow conditions (THF) for both methods and better column efficiencies are also exhibited towards PAHs' separation compared to the classical coated stationary phase. The characterization of these new stationary phases and the rationale for improved column stability have been investigated by solid state 13C and 29Si CP/MAS NMR spectroscopy. It is clearly shown that the chemical bonding is achieved by the classical hydrosilylation reaction between PHMS chains and vinyl modified silica. The bonded polymer is likely a copolymer than a homopolymer. The immobilization of the SC-LCP by heating results in the breaking of Si-O-Si bonds of the polysiloxane chain after the attack of the silica surface silanols. Applications to fullerenes and carotenes separation of these bonded stationary phases are compared to the separation power of a classical monomeric C18 stationary phase in NP-HPLC as n-hexane-toluene or methyl-tertiobutyl ether-methanol mixtures.