Photoregulation of Gene Expression with Ligand-Modified Caged siRNAs through Host/Guest Interaction

Small interfering RNA (siRNA) can effectively silence target genes through Argonate 2 (Ago2)-induced RNA interference (RNAi). It is very important to control siRNA activity in both spatial and temporal modes. Among different masking strategies, photocaging can be used to regulate gene expression through light irradiation with spatiotemporal and dose-dependent resolution. Many different caging strategies and caging groups have been reported for light-activated siRNA gene silencing. Herein, we describe a novel caging strategy that increases the blocking effect of RISC complex formation/process through host/guest (including ligand/receptor) interactions, thereby enhancing the inhibition of caged siRNA activity until light activation. This strategy can be used as a general approach to design caged siRNAs for the photomodulation of gene silencing of exogenous and endogenous genes.     

Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

纳米SiO_2与有机物分子的亲和性和分散性

介绍纳米SiO2的合成和粒子表面的粗糙形貌以及纳米粒子链具有类似高分子链的弹性力学属性,研究纳米SiO2的亲合性,探讨亲合性对纳米SiO2的分散、改性、应用的影响。研究表明,在极性外加剂(如乙二胺、乙二醇、PEG、PDMS、PTHF、PMMA、PBMA等)存在的情况下,纳米SiO2的弹性粒子链和硅烷醇对SiO2的分散和改性具有显著的影响。带有大烷基侧链的高分子链与SiO2表面的亲合能较低,而带有官能团的高分子链与SiO2表面具有较高的亲合性。     

Isolation of the acidic and basic lectins from winged bean seed (Psophocarpus tetragonolobus (L.) DC)

The acidic and basic lectins of the winged bean were isolated from seed extracts by affinity chromatography on lactose-Sepharose 6B and melibiose Bio-Gel P150, respectively. The lectins isolated from six varieties of Psophocarpus tetragonolobus, collected from different regions of south-east Asia, showed no significant differences in their properties; their subunit M_r and amino acid compositions were the same as those of the acidic and basic lectins isolated by non-affinity methods. On isoelectric focusing the affinity purified acidic lectins showed two major protein components (isolectins) while the affinity purified basic lectins had four protein components. The acidic lectins showed a preference for β-D-galactosides and they agglutinated trypsinised human type A, B and O erythrocytes but not trypsinised rabbit erythrocytes. The basic lectins showed a preference for α-D-galactosides and they agglutinated trypsinised rabbit and human type A and B erythrocytes but not trypsinised human type O erythrocytes.     

Enantioselective extraction of mandelic acid enantiomers based on chiral ligand exchange

Based on the chiral ligand exchange, the distribution behavior of mandelic acid enantiomers, and the partition of Cu^2 at different pH values were studied in a water/alcohol two-phase system containing Cu^2 and N-ndodecyl-L-proline(A). The influences of the solvent sort, the pH value, the concentrations of Cu^2 and chiral ligand on the partition coefficient(K) and separation factor(α) were discussed. The experimental results show that the A formed has more stable ternary complex with D-mandelic acid enantiomer than with L-mandelic acid enantiomer. There is an important influence of the pH value on K and a. When the pH values are less than 3.5, the formation of binary complexes is thermodynamically unfavourable. K and a become maximum when pH values are above 3.5 and the molar ratio of the chiral ligand to Cu^2 is 2:1.     

Thermodynamic state of reinforced interpenetrating polymer networks

The free energies of mixing of two networks in the interpenetrating polymer network based on crosslinked polyurethane and poly(ester acrylate) have been determined by the vapour sorption method. It was established that the constituent networks in the IPN are not miscible. The introduction of fillers of different chemical nature increases the compatibility. The thermodynamic affinity of the fillers to the individual networks and IPN was estimated. It was established that when the free energy of interaction of one or both components of the IPN with the filler is negative, reinforcement leads to the formation of a compatible and equilibrium system. For fillers having no affinity to the polymers, compatibilization is observed, which is connected with slowing down of phase separation in the system in the presence of filler.     

Synthesis,Characterization and Luminescent Properties of Complexes of Rare Earth Nitrates with Tris{2-[N-(pyridine-methanoyl)amino]ethyl} amine

Duringthelastdecade ,thedesignandsynthesisoftripodalligandsandtheirmetalcomplexeshaveattractedconsiderableattentionduetotheirpotentialusesaslumi nescentprobesinmedicineandbiochemistry[1～ 7] ,ascatalysts[8] andelectrochemicalmaterials[9] .However ,thecomplexesofrareearthmetalwithtripodalligandshavefewreported[10 ] .Asanextensionofourstudiesoncomplexesofmetalwithtripodalligands ,anewtripodalligand (tris { 2 [N (pyridine methanoyl)amino]ethyl}amine ,L)anditscomplexeswithrareearthnitratesweresyn…     

Rational Design of CYP3A4 Inhibitors: A One-Atom Linker Elongation in Ritonavir-Like Compounds Leads to a Marked Improvement in the Binding Strength

Inhibition of the major human drug-metabolizing cytochrome P450 3A4 (CYP3A4) by pharmaceuticals and other xenobiotics could lead to toxicity, drug–drug interactions and other adverse effects, as well as pharmacoenhancement. Despite serious clinical implications, the structural basis and attributes required for the potent inhibition of CYP3A4 remain to be established. We utilized a rational inhibitor design to investigate the structure–activity relationships in the analogues of ritonavir, the most potent CYP3A4 inhibitor in clinical use. This study elucidated the optimal length of the head-group spacer using eleven (series V) analogues with the R₁/R₂ side-groups as phenyls or R₁–phenyl/R₂–indole/naphthalene in various stereo configurations. Spectral, functional and structural characterization of the inhibitory complexes showed that a one-atom head-group linker elongation, from pyridyl–ethyl to pyridyl–propyl, was beneficial and markedly improved K_s, IC₅₀ and thermostability of CYP3A4. In contrast, a two-atom linker extension led to a multi-fold decrease in the binding and inhibitory strength, possibly due to spatial and/or conformational constraints. The lead compound, 3h, was among the best inhibitors designed so far and overall, the strongest binder (K_s and IC₅₀ of 0.007 and 0.090 µM, respectively). 3h was the fourth structurally simpler inhibitor superior to ritonavir, which further demonstrates the power of our approach.     

Accurate Receptor-Ligand Binding Free Energies from Fast QM Conformational Chemical Space Sampling

Small molecule receptor-binding is dominated by weak, non-covalent interactions such as van-der-Waals hydrogen bonding or electrostatics. Calculating these non-covalent ligand-receptor interactions is a challenge to computational means in terms of accuracy and efficacy since the ligand may bind in a number of thermally accessible conformations. The conformational rotamer ensemble sampling tool (CREST) uses an iterative scheme to efficiently sample the conformational space and calculates energies using the semi-empirical ‘Geometry, Frequency, Noncovalent, eXtended Tight Binding’ (GFN2-xTB) method. This combined approach is applied to blind predictions of the modes and free energies of binding for a set of 10 drug molecule ligands to the cucurbit[n]urils CB[8] receptor from the recent ‘Statistical Assessment of the Modeling of Proteins and Ligands’ (SAMPL) challenge including morphine, hydromorphine, cocaine, fentanyl, and ketamine. For each system, the conformational space was sufficiently sampled for the free ligand and the ligand-receptor complexes using the quantum chemical Hamiltonian. A multitude of structures makes up the final conformer-rotamer ensemble, for which then free energies of binding are calculated. For those large and complex molecules, the results are in good agreement with experimental values with a mean error of 3 kcal/mol. The GFN2-xTB energies of binding are validated by advanced density functional theory calculations and found to be in good agreement. The efficacy of the automated QM sampling workflow allows the extension towards other complex molecular interaction scenarios.