Molecular imprinted Nylon-6 as a recognition material of amino acids

Nylon-6 was functionalized by phase inversion molecular imprinting technique to introduce l-glutamine recognition property. For imprinting l-glutamine in the polymer, 20 wt.% of Nylon-6-formic acid solution with 8 wt.% of l-glutamine template was used for the phase inversion process in water. The resulted polymer including the template molecule was washed with acetic acid solution to extract the template from the polymer. The substrate binding experiments of the l-glutamine imprinted polymer were examined in aqueous l-glutamine, d-glutamine, l-glutamic acid, and d-glutamic acid solution. The binding of l-glutamine increased with the increase of the amino acid concentration from 5 to 20 μM. The value of equilibrium binding constant for l-glutamine, K_E=4.9×10⁵ M⁻¹, was larger than that for d-glutamine, K_E=1.5×10⁵ M⁻¹. The recognition experiments were extended to membrane filtration and quartz-crystal microbalance response by using the imprinted Nylon-6. Evidence was also presented by FT-IR analysis that the amide-hydrogen-bonding interaction between the imprinted Nylon-6 and template was originated for the amino acid recognition.     

Enantioselective CD analysis of amino acids based on chiral amplification with a stereodynamic probe

The condensation between stereolabile 1,8-bis(3′-formyl-4′-hydroxyphenyl)naphthalene, 1, and two amino acid molecules results in the formation of chiral diimines exhibiting strong CD signals. This reaction has been used to develop a chiroptical sensing method for the determination of the absolute configuration and enantiomeric composition of unprotected amino acids. This sensing approach is based on distinctive chiral amplification due to central-to-axial chirality induction within the diimine scaffold formed and does not require the use of an enantiopure ligand or metal complex.     

The use of acridine orange base (AOB) as molecular probe to characterize nonaqueous AOT reverse micelles

The behavior of acridine orange base (AOB) in nonaqueous reverse micelles composed of n-heptane/AOT/polar solvent has been performed. Ethylene glycol (EG), propylene glycol (PG), glycerol (GY), formamide (FA), dimethylformamide (DMF), and dimethylacetamide (DMA) were employed as water substitutes. The studies were performed by static and time-resolved emission spectroscopy. Thus, the distribution of AOB between the two pseudophases of the aggregates was quantified by measuring the partition constants from emission spectra at different surfactant concentration. Similar values to those obtained by means of absorption spectroscopy were obtained. This match is indicating that AOB is not experiencing partition during the lifetime of the excited state. Partitioning to the micelles is strongly favored in micelles containing hydrogen-bond donor (HBD) solvents rather than non-HBD solvents. Variations of fluorescence lifetimes with AOT concentration confirm these results. By the solvatochromic behavior of AOB in the different systems it is shown that the microenvironment at the interface is distinct from that of the bulk polar solvent, indicating that the probe senses no "free" solvent. The steady state anisotropy (r) was measured for EG/AOT/n-heptane and DMF/AOT/n-heptane systems as representatives for HBD and non-HBD polar solvents, respectively. The value of r is higher in the micelles containing EG than that obtained with DMF, and increases with AOT concentration. This is explained as due to highly structured polar solvents in the inner core. EG is interacting with the polar heads of AOT through hydrogen-bond interaction, while DMF can only interact with the Na+ counterions. This is confirmed by the time-resolved emission spectra (TRES) of the probe in the micellar systems, in comparison with the bulk solvents.     

Pyrene-appended alpha-cyclodextrin as a fluorescent pH probe responding to a wide range.

Pyrene-appended alpha-cyclodextrin (3) in which a trimethylenediamine linker connected the pyrene residue to the alpha-cyclodextrin moiety showed pH-dependent fluorescence intensity changes. The fluorescence intensity was almost linearly changed within the pH range of 5 - 10. The unique fluorescence response of 3 to the pH was due not only to the favorable pK(a) values (pK(a1) = 6.4 and pK(a2) = 8.8), but also to the almost equal contributions of the amino groups to the pyrene's fluorescence quenching.     

Molecular motions of alpha-cyclodextrin on a dodecyl chain studied by molecular dynamics simulations

Motions of an alpha-cyclodextrin (alpha-CD) molecule on a dodecyl chain adopting the all-trans conformation were investigated in the presence of water by molecular dynamics simulations with CVFF force fields, where the trimethylammonium group of dodecyltrimethylammonium bromide (DTAB) is protruded outside the secondary hydroxyl rim of alpha-CD (the secondary-in structure). The alpha-CD molecule shuttled rapidly on the chain without decomplexation. This rapid motion is consistent with the NMR data. The plane formed by 6 O4 atoms of alpha-CD is most populated between the C6 and C7 atoms of DTAB. This structure is very close to that estimated by NMR. The alpha-CD molecule underwent a restricted rotation in a range of 60 degrees with regard to the plane of the dodecyl chain: this plane at the most population is middle between the two diagonal lines of the normal hexagon formed by 6 O4 atoms of alpha-CD. The published NMR data were reanalyzed in terms of the rotation angle, and a slightly better structure was obtained. The distortion of the alpha-CD cavity from the normal hexagon was decreased upon complex formation with DTAB. The deviation of the center of alpha-CD from the center of the dodecyl chain predicted by molecular dynamics simulations is consistent with the NMR data. The secondary-in structure is energetically more stable than the primary-in structure, as calculated by molecular mechanics with CVFF and Amber force fields. This result is consistent with the NMR data. Molecular dynamics simulations were also carried out for the primary-in structure. Some of the results are close to those of the secondary-in structure.     

Analysis of amino acids and proteins using a poly(methyl methacrylate) microfluidic system

Plastic microchips are very promising analytical devices for the high-speed analysis of biological compounds. However, due to its hydrophobicity, their surface strongly interacts with nonpolar analytes or species containing hydrophobic domains, resulting in a significant uncontrolled adsorption on the channel walls. This paper describes the migration of fluorescence-labeled amino acids and proteins using the poly(methyl methacrylate) microchip. A cationic starch derivative significantly decreases the adsorption of analytes on the channel walls. The migration time of the analytes was related to their molecular weight and net charge or pI of the analytes. FITC-BSA migrated within 2 min, and the theoretical plate number of the peak reached 480,000 plates/m. Furthermore, proteins with a wide range of pI values and molecular weights migrated within 1 min using the microchip.     

Molecular recognition of nucleobases and amino acids by sulphonato-calixnaphthalene-capped silver nanoparticles

The plasmon resonances of sulphonato-calixnaphthalene-capped silver nanoparticles have been used to study the complexation of the nanoparticles with nucleobases and amino acids. Only in the case of the nanoparticles capped with oxacalix[4]naphthalenesultone, does complexation of both nucleobases and certain amino acids occur. The complexation of the aromatic amino acids, phenylalanine and tryptophan, has previously not been observed for calixarene-capped silver nanoparticles.     

Thiazole orange (TO) as a light-switch probe: a combined quantum-mechanical and spectroscopic study

A Density Functional Theory (DFT) study of the absorbance and fluorescence emission characteristics of the cyanine thiazole orange (TO) in solution and when intercalated in DNA was carried out in combination with spectrophotometric and spectrofluorometric experiments under different conditions (temperature, concentration, solvent viscosity). T-jump relaxation kinetics of the TO monomer-dimer conversion enabled the thermodynamic parameters of this process to be evaluated. The overall data collected provided information on the features of the "light-switch" by the fluorescent TO and the comparison between experimental and calculated photo-physical properties allowed us to explain and rationalize both shifts and quenching/enhancing effects on fluorescence due to solvation, dimerisation and intercalation in the DNA.     

Synthesis of nonproteinogenic amino acids to probe lantibiotic biosynthesis

The synthesis of six nonproteinogenic amino acids appropriately protected for Fmoc-based solid-phase peptide synthesis is described. These amino acids are (2S,3R)-vinylthreonine, (2S)-(E)-2-amino-5-fluoro-pent-3-enoic acid (fluoroallylglycine), (S)-beta(2)-homoserine, (S) and (R)-beta(3)-homocysteine, and (2R,3R)-methylcysteine. Once incorporated into peptides, these compounds were envisioned to serve as alternative substrates for the lantibiotic synthases that dehydrate serine and threonine residues in their peptide substrates and catalyze the subsequent intramolecular Michael-type addition of cysteines to the dehydroamino acids.     

The foundation of a light driven molecular muscle based on stilbene and alpha-cyclodextrin

The rotaxane serves as the basis of a light driven molecular muscle, where reversible photoisomerisation of the stilbene units causes the cyclodextrins to move off and on the stilbene units, contracting and extending the distance between the blocking groups.     

Polyamides of 2,5-bis(amino methyl) 1,4-dioxane and dicarboxylic acids

Novel polyamides of 2,5-bis(amino methyl) 1,4-dioxane (cis/trans-BAMD) with adipic/sebacic acids and of 2,5-bis(carboxy methyl)/2,5-bis(carboxy) 1,4-dioxane with 1,6-hexane diamine have been prepared. Because of the slightly higher conformational flexibility of the 1,4-dioxane ring in comparison with that of the cyclohexane ring, the BAMD polyamides have lower T_g and T_m than the corresponding 1,4-bis(amino methyl) cyclohexane (BAMC) polyamides. The symmetry of the trans-dioxane moiety permits high crystallinity in the trans-BAMD polymers. The crystallinity T_g and T_m of the trans polymers are decreased with the incorporation of the cis-dioxane moiety which lacks a plane of symmetry. Because of the hydrophilic nature of the dioxane ring, t-BAMD-6 has good moisture-regain properties, yet is melt processable (T_m = 292°C).     

Molecular recognition properties of acyclic cucurbiturils toward amino acids,peptides, and a protein

The binding of 1 and 2 toward 19 amino acid amides by ¹H NMR and ITC is reported. Hosts 1 and 2 bind to aromatic or hydrophobic residues by cavity inclusion leaving the cationic residues at the C=O portals. K_a values range from 10² to >10⁶ M^?1 with H-Phe-NH₂, H-Trp-NH₂, and H-Tyr-NH₂ displaying sub-micromolar K_d values. Hosts 1 and 2 bind tightly to dicationic H-Lys-NH₂ and H-Arg-NH₂ which are poor guests for CB[7]. Comparison of the affinity of 1 and 2 toward the amino acid amide, N-acetyl-amino-acid amide, and amino acid forms of Phe revealed that the removal of the NH₃⁺ to O=C and SO₃^? electrostatic interactions costs 3.8 kcal/mol whereas the introduction of an unfavourable CO₂^? to O=C and SO₃^? electrostatic interactions costs 2.1 kcal/mol. Hosts 1 and 2 bind to insulin with low micromolar affinity. Acyclic CB[n] display high affinity toward a wider range of N-terminal amino acids residues than CB[n] which suggests a broad range of applications.     

A novel histidine assay using tetraphenylporphyrin manganese (III) chloride as a molecular recognition probe by resonance light scattering technique

A novel histidine-selective method has been developed for the determination of histidine in aqueous solutions by resonance light scattering (RLS) technique. At pH 8.0, the weak RLS intensity of tetraphenylporphyrin manganese (III) chloride [MnTPPCl] was greatly enhanced by the addition of histidine with the maximum peak located at 483 nm. Under the optimum conditions, it was found that the enhanced RLS intensity was in proportion to the concentration of histidine in the range 7.8 × 10⁻⁷-2.4 × 10⁻⁵ mol l⁻¹. Low detection limit of 9.2 × 10^-8 mol l⁻¹ has been achieved. The histidine concentrations in synthetic samples and real samples were determined with satisfactory results. The sensitivity and selectivity of this method are high enough to permit the determination of trace amounts of histidine without any significant interference from high levels of other components such as common anions and especially, other amino acids.     

Synthesis of poly(ethylene glycol methyl ether)-b-poly(ethylenimine) and its derivatives containing thymine and amino acids as pendants

Poly(ethylene glycol methyl ether)tosylate was prepared and used to initiate the polymerization of 2-methyl-2-oxazoline. The resulting poly(ethylene glycol methyl ether)-b-poly(N-acetyl ethylenimine) was hydrolyzed and neutralized to give poly(ethylene glycol methyl ether)-b-poly(ethyl-enimine) (PEO–PEI). 2-(thymin-1-yl)propionic acid, N-Cbz-alanine, N-Cbz-proline, N-Cbz-O-t-Bu-serine. and N-FMOC-proline were grafted onto the PEO–PEI copolymer; attempts were then made to remove the Cbz and FMOC protecting groups.     

Incorporation of extra amino acids in peptide recognition probe to improve specificity and selectivity of an electrochemical peptide-based sensor

We investigated the effect of incorporating extra amino acids (AA) at the n-terminus of the thiolated and methylene blue-modified peptide probe on both specificity and selectivity of an electrochemical peptide-based (E-PB) HIV sensor. The addition of a flexible (SG)₃ hexapeptide is, in particular, useful in improving sensor selectivity, whereas the addition of a highly hydrophilic (EK)₃ hexapeptide has shown to be effective in enhancing sensor specificity. Overall, both E-PB sensors fabricated using peptide probes with the added AA (SG-EAA and EK-EAA) showed better specificity and selectivity, especially when compared to the sensor fabricated using a peptide probe without the extra AA (EAA). For example, the selectivity factor recorded in the 50% saliva was ∼2.5 for the EAA sensor, whereas the selectivity factor was 7.8 for both the SG-EAA and EK-EAA sensors. Other sensor properties such as the limit of detection and dynamic range were minimally affected by the addition of the six AA sequence. The limit of detection was 0.5 nM for the EAA sensor and 1 nM for both SG-EAA and EK-EAA sensors. The saturation target concentration was ∼200 nM for all three sensors. Unlike previously reported E-PB HIV sensors, the peptide probe functions as both the recognition element and antifouling passivating agent; this modification eliminates the need to include an additional antifouling diluent, which simplifies the sensor design and fabrication protocol.     

A cyclometalated palladium-azo complex as a differential chromogenic probe for amino acids in aqueous solution

Solutions of a cyclometalated palladium-azo complex exhibited differential UV-Vis absorption spectra in the presence of alpha-amino acids with different side chain groups.     

A novel approach to the synthesis of amino acids based on cobalt bis(dicarbollide)

A novel approach to the synthesis of boron-containing amino acids based on ring opening of cyclic oxonium derivatives of polyhedral boron hydrides under the action of the terminal functional groups of natural amino acids was proposed. This approach was successfully implemented for the synthesis of cobalt bis(dicarbollide) — tyrosine conjugate.     

二聚炔雌醇分子钳对氨基酸甲酯手性识别的理论研究

采用分子模拟研究了二聚炔雌醇分子钳(1~3)与D-和L-苯丙氨酸甲酯的相互作用,比较了形成配合物前后分子钳的结构变化以及与手性苯丙氨基酸甲酯相互作用能大小,其计算结果与实验结果一致。理论研究表明主客体分子间存在着的空间匹配、Van der Waals作用力和静电作用是分子识别的推动力,这为认识和预测分子钳的手性识别能力提供了理论依据。     

Binding of calcium to amino acids (2)

Crystal structures ofN-acetyl-(DL)-methionine and its calcium adduct have been determined in order to examine possible calcium coordination through sulfur atom and conformation changes in the molecule upon calcium coordination. Crystals of the Ca adduct ofN-acetyl-(DL)-methionine are triclinic, space groupP¯1, a=11.356(2) Å,b=13.556(2) Å,c=15.035(3) Å,=95.42(1)°,=103.66(1)°, and=103.66(1)°, withZ=4 andd _o= 1.40 g cm^–3; 5834 reflections were collected at –125°C. TheN-acetyl-(DL)-methionine was also crystallized so that changes induced by the calcium coordination could be observed. TheN-acetyl-(DL)-methionine crystals are monoclinic, space groupP2₁/c,a=5.885(1) Å,b=9.290(1) Å,c=18.024(2) Å,=92.51 (1)°, withZ=4 andd _o=1.30 g cm^–3. Both structures were solved using direct methods and refined using full matrix least squares. The final values ofR andR _w were 0.051 and 0.053 for the Ca adduct and 0.048 and 0.039 for the free amino acid. All hydrogen atoms were located and included in the final calculations. Interatomic distances and angles of the four crystallographically independentN-acetylmethionine molecules in the Ca adduct are comparable to those of the free molecule, but the torsion angles change drastically. Calcium coordination through the sulfur atom is not found. The binding modes of all four independentN-acetylmethionine molecules in the Ca adduct are different. Of the two crystallographically independent calcium atoms one is 6 and the other is 7 coordinated. The calcium adduct shows several Ca-Ca bridges (with distances 4.722, 4.746, 5.503, and 6.265 Å) which can be described as a zigzag calcium chain or alternatively as slightly puckered sheets of calcium atoms parallel to theab plane atz O.     

氨基甲酸酯型脱氧胆酸分子钳对氨基酸甲酯的手性识别 研究

以脱氧胆酸为spacer,通过三光气桥连各种芳胺,合成了新的氨基甲酸酯型分子钳受体1～4,这些化合物的结构经IR,^1HNMR和元素分析所证实。利用差光谱滴定法考察了其与D/L氨基酸甲酯的对映选择性识别性能。结果表明,分子钳1～4对所考察的氨基酸甲酯均具有识别能力,其对D-氨基酸甲酯的识别优于对L-氨基酸甲酯的识别。从主客体间的大小形状匹配及几何互补关系等方面对这些受体的识别能力及对映选择性进行了讨论。