Differential ordering of the protein backbone and side chains during protein folding revealed by site-specific recombinant infrared probes

The time scale for ordering of the polypeptide backbone relative to the side chains is a critical issue in protein folding. The interplay between ordering of the backbone and ordering of the side chains is particularly important for the formation of β-sheet structures, as the polypeptide chain searches for the native stabilizing cross-strand interactions. We have studied these issues in the N-terminal domain of protein L9 (NTL9), a model protein with mixed α/β structure. We have developed a general approach for introducing site-specific IR probes for the side chains (azide) and backbone ((13)C═(18)O) using recombinant protein expression. Temperature-jump time-resolved IR spectroscopy combined with site-specific labeling enables independent measurement of the respective backbone and side-chain dynamics with single residue resolution. We have found that side-chain ordering in a key region of the β-sheet structure occurs on a slower time scale than ordering of the backbone during the folding of NTL9, likely as a result of the transient formation of non-native side-chain interactions.     

Role of parallel reformable bonds in the self-healing of cross-linked nanogel particles

We develop a hybrid computational approach to examine the mechanical properties and self-healing behavior of nanogel particles that are cross-linked by both stable and labile bonds. The individual nanogels are modeled via the lattice spring model (LSM), which is an effective method for probing the response of materials to mechanical deformation. The cross-links between the nanogels are simulated via the hierarchical Bell model (HBM), which allows us to capture the rupturing of multiple parallel bonds as the result of an applied force. Because the labile bonds are relatively reactive, they can reform after they have been ruptured. To incorporate the possibility of bonds reforming, we modify the HBM formalism and validate the modified HBM by considering a system of two surfaces, which are connected by multiple parallel bonds. We then use our hybrid HBM/LSM to simulate the behavior of the cross-linked nanogels under a tensile deformation. In these simulations, each labile linkage between the nanogels contains at most N parallel bonds. We vary the fraction of labile linkages and the value of N in these linkages to determine the optimal conditions for improving the robustness of the material. Although numerous parallel bonds within a linkage enhance the strength of the material, these bonds diminish the ductility and the ability of the material to undergo the structural rearrangements that are necessary for self-repair. For a relatively low fraction of labile bonds and N ≤ 4, however, we can significantly improve the strength of the material and preserve the self-healing properties. For instance, a sample with 30% labile linkages and N = 4 per linkage is roughly 200% stronger than a sample that is cross-linked solely by stable bonds and can still undergo self-repair in response to the tensile deformation. The results reveal how mechanical stress can lead not only to the appearance of cavities within the material but also to bond formation that "heals" these cavities and thus prevents the catastrophic failure of the material.     

Analysis of drug interactions with serum proteins and related binding agents by affinity capillary electrophoresis: A review

Biomolecules such as serum proteins can interact with drugs in the body and influence their pharmaceutical effects. Specific and precise methods that analyze these interactions are critical for drug development or monitoring and for diagnostic purposes. Affinity capillary electrophoresis (ACE) is one technique that can be used to examine the binding between drugs and serum proteins, or other agents found in serum or blood. This article will review the basic principles of ACE, along with related affinity-based capillary electrophoresis (CE) methods, and examine recent developments that have occurred in this field as related to the characterization of drug–protein interactions. An overview will be given of the various formats that can be used in ACE and CE for such work, including the relative advantages or weaknesses of each approach. Various applications of ACE and affinity-based CE methods for the analysis of drug interactions with serum proteins and other binding agents will also be presented. Applications of ACE and related techniques that will be discussed include drug interaction studies with serum agents, chiral drug separations employing serum proteins, and the use of CE in hybrid methods to characterize drug binding with serum proteins.     

Mononuclear and Dinuclear Copper Complexes of Tridentate Redox-active Ligands with Tunable H-bonding Donors: Structure,Spectroscopy and H+/e− Reactivity

In this research article, we describe the synthesis and characterization of mononuclear and dinuclear Cu complexes bound by a family of tridentate redox-active ligands with tunable H-bonding donors. The mononuclear Cu-anion complexes were oxidized to the corresponding “high-valent” intermediates by oxidation of the redox-active ligand. These species were capable of oxidizing phenols with weak O−H bonds via H-atom abstraction. Thermodynamic analysis of the H-atom abstractions, which included reduction potential measurements, pK_a determination and kinetic studies, revealed that modification of the anion coordinated to the Cu and changes in the H-bonding donor did not lead to major differences in the reactivity of the “high-valent” CuY complexes (Y: hydroxide, phenolate and acetate), which indicated that the tridentate ligand scaffold acts as the H⁺ and e⁻ acceptor.     

Dynamic Ion Speciation during the Hydrolysis of Aryltrifluoroborates**

Organotrifluoroborates serve as coupling partners during transmetalation in the Suzuki–Miyaura reaction but require hydrolysis prior to the coupling reaction. Their anionic nature allows study of their hydrolysis by electrospray ionization mass spectrometry (ESI-MS) through real-time monitoring, complemented by pH analysis. The induction period varied according to the borates employed, and a dynamic series of equilibria for numerous ions was observed during hydrolysis. We found that the induction periods and reaction rates were sensitive to the R group of the borates, the shape of the reaction vessel, and stir rate.     

[Ni(NHC)2] as a Scaffold for Structurally Characterized trans [H−Ni−PR2] and trans [R2P−Ni−PR2] Complexes

The addition of PPh₂H, PPhMeH, PPhH₂, P(para-Tol)H₂, PMesH₂ and PH₃ to the two-coordinate Ni⁰ N-heterocyclic carbene species [Ni(NHC)₂] (NHC=IiPr₂, IMe₄, IEt₂Me₂) affords a series of mononuclear, terminal phosphido nickel complexes. Structural characterisation of nine of these compounds shows that they have unusual trans [H−Ni−PR₂] or novel trans [R₂P−Ni−PR₂] geometries. The bis-phosphido complexes are more accessible when smaller NHCs (IMe₄>IEt₂Me₂>IiPr₂) and phosphines are employed. P−P activation of the diphosphines R₂P−PR₂ (R₂=Ph₂, PhMe) provides an alternative route to some of the [Ni(NHC)₂(PR₂)₂] complexes. DFT calculations capture these trends with P−H bond activation proceeding from unconventional phosphine adducts in which the H substituent bridges the Ni−P bond. P−P bond activation from [Ni(NHC)₂(Ph₂P−PPh₂)] adducts proceeds with computed barriers below 10 kcal mol⁻¹. The ability of the [Ni(NHC)₂] moiety to afford isolable terminal phosphido products reflects the stability of the Ni−NHC bond that prevents ligand dissociation and onward reaction.     

Strawberry Juice Powders: Effect of Spray-Drying Conditions on the Microencapsulation of Bioactive Components and Physicochemical Properties

The drying of fruit juices has advantages such as easy handling of powders, reduction in volume, and preservation of the characteristics of the fruit. Thus, in this work, the effect of the spray drying conditions of strawberry juice (SJ) with maltodextrin (MX) as a carrying agent on the microencapsulation of bioactive compounds and physicochemical properties was studied. The content of phenolic compounds and antioxidant activity showed higher values at low concentrations of MX, while the effect of drying temperature was negligible. The thermal characterization showed that the low molecular weight sugars in the juice decreased the glass transition temperature (Tg). The morphological analysis by scanning electron microscopy (SEM) indicated that at low concentrations of MX, the particles agglomerated, while at intermediate and high concentrations, the particles were observed as well separated. Through microstructural analysis by X-ray diffraction (XRD), the presence of amorphous state was confirmed in all the samples, which is beneficial for preventing chemical and biochemical reactions, and promoting the conservation of the microencapsulated bioactive compounds.     

Study of the chromonic liquid-crystalline phases of bis-(N,N-diethylaminoethyl)perylene-3,4,9,10-tetracarboxylic diimide dihydrochloride by polarized optical microscopy and 2H NMR spectroscopy

The chromonic liquid-crystalline properties of bis-(N,N-diethylaminoethyl)perylene-3,4,9,10-tetracarboxylic diimide dihydrochloride in an aqueous solution were investigated by polarized light microscopy and 2H NMR spectroscopy. Both techniques indicate a narrow I + N biphasic region and a broad N phase region at concentrations ranging from approximately 6.9 to approximately 30 wt % at room temperature. Optical microscopy indicates that a hexagonal M phase exists at higher concentrations. The variation of the I --> N + I and N + I --> N transition temperatures with concentration was studied by 2H NMR spectroscopy. Finally, the effects of temperature and concentration on the order parameter of the N phase were investigated by 2H NMR using a tetra-deuterated derivative. A value of 0.97 was obtained for the N phase at its upper concentration limit.