The development of the Diels-Alder cycloaddition as a new method for the site-specific chemoselective ligation of peptides and proteins under mild conditions is reported. Peptides equipped with a 2,4-hexadienyl ester and an N-terminal maleimide react in aqueous media to give cycloadducts in high yields and depending on the amino acid sequence with high stereoselectivity. Except for the cysteine SH group the transformation is compatible with all amino acid side chain functional groups. For ligation to proteins the hexadienyl group was attached to avidin and streptavidin noncovalently by means of complex formation with a biotinylated peptide or by covalent attachment of a hexadienyl ester-containing label to lysine side chains incorporated into the proteins. Site-specific attachment of the hexadienyl unit into a Rab protein was achieved by means of expressed protein ligation followed by protection of the generated cysteine SH by means of Ellman's reagent. The protein reacted with different maleimido-modified peptides under mild conditions to give the fully functional cycloadducts in high yield. The results demonstrate that the Diels-Alder ligation offers an advantageous and technically straightforward new opportunity for the site-specific equipment of peptides and proteins with further functional groups and labels. It proceeds under very mild conditions and is compatible with most functional groups found in proteins. Its combination with other ligation methods, in particular expressed protein ligation is feasible. 相似文献
A series of epoxy cured samples, with different molar composition of 1,4-butanediol diglycidyl ether (EP), cis-1,2-cyclohexanedicarboxylic anhydride (CH) and triethylamine (TEA) as initiator, was studied by Electrical Impedance Spectroscopy (EIS) in the frequency range 10−2-105 Hz. The resistivity of the system was evaluated by using the complex impedance analysis method. Resistivity values in the range between 107 and 1013 Ω cm were obtained and related to the composition (molar fraction xEP or xCH) of the epoxy system. The gel formation characteristics of the samples were obtained as theoretically described by Flory. 相似文献
Samples of must derived from six different varieties of grapes taken during the fermentation process, as well as the respective wine samples directly after the end of the malolactic fermentation, were analyzed by direct infusion negative ion mode electrospray ionization mass spectrometry (ESI-MS). Diagnostic ions for must were different from those of wine samples, although small variations for each of the grape varieties were also detected. The addition of unfermented must or sugar to wine could also be clearly detected. The spectra were acquired in a few minutes per sample, indicating that ESI-MS can be used for high-throughput analysis of samples and should prove useful for quality control during and after the fermentation process. 相似文献
Summary: Two peptides (FEFEFKFK and HHHHHHFEFEFKFK) have been synthesised and their phase diagrams mapped out as a function of concentration and temperature. Both peptides formed self-supporting fibrillar hydrogels above a similar critical molar gelation concentration with a fibril diameter corresponding to the length of the fully stretched monomer. Mixing the peptides in a 9:1 ratio of FEFEFKFK to histadine functionalized FEFEFKFK in the presence of nanogold particles (binds to the histadine groups) resulted in thicker fibres suggesting that two fibrils associate together to form a fibre. TEM studies revealed that the gold particles were distributed throughout the hydrogel and adjacent to both sides of the fibrillar structures with an average distance between particles of 21 nm. It is postulated that the peptides form anti-parallel beta sheet fibrils that associate together via π-stacking interactions between the imidazole side chains of the histadine groups to form a fibre, where on average 1 in every 44 peptides is functionalized. 相似文献
Enzymes are attractive, “green” alternatives to chemical catalysts within the industrial sector, but their robustness to environmental conditions needs optimizing. Here, an enzyme is tagged chemically and recombinantly with a self‐assembling peptide that allows the conjugate to spontaneously assemble with pure peptide to form β‐sheet‐rich nanofibers decorated with tethered enzyme. Above a critical concentration, these fibers entangle and form a 3D hydrogel. The immobilized enzyme catalyzes chemical transformations and critically its stability is increased significantly where it retains activity after exposure to high temperatures (90 °C) and long storage times (up to 12 months).
The antimicrobial and cytotoxic activities of the dichloromethane, ethyl acetate and butanolic fractions from the leaves, twigs and stem bark of Scutia buxifolia were evaluated using the broth microdilution method and the brine shrimp lethality method, respectively. Phytochemical analysis was performed by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The antimicrobial results demonstrated that the strongest effect occurred with the butanol fraction from the twigs and the ethyl acetate fraction from the stem bark against Saccharomyces cerevisiae (minimal inhibitory concentration; MIC?=?62.5?μg?mL(-1)), whereas the ethyl acetate and butanolic fractions from the twigs and stem bark were effective against Pseudomonas aeruginosa and Staphylococcus aureus, with MIC values ranging from 125 to 500?μg?mL(-1). LD(50) values varied from 50.00?±?0.22 to 82.23?±?0.34?μg?mL(-1). Quercetin, quercitrin, isoquercitrin and rutin were identified by HPLC and may be partially responsible for the antimicrobial activities observed. This study reports for the first time the antimicrobial and cytotoxic activities of S. buxifolia leaves, twigs and stem bark. 相似文献
As potential inhibitors of penicillin‐binding proteins (PBPs), we focused our research on the synthesis of non‐traditional 1,3‐bridged β‐lactams embedded into macrocycles. We synthesized 12‐ to 22‐membered bicyclic β‐lactams by the ring‐closing metathesis (RCM) of bis‐ω‐alkenyl‐3(S)‐aminoazetidinone precursors. The reactivity of 1,3‐bridged β‐lactams was estimated by the determination of the energy barrier of a concerted nucleophilic attack and lactam ring‐opening process by using ab initio calculations. The results predicted that 16‐membered cycles should be more reactive. Biochemical evaluations against R39 DD‐peptidase and two resistant PBPs, namely, PBP2a and PBP5, revealed the inhibition effect of compound 4d , which featured a 16‐membered bridge and the N‐tert‐butyloxycarbonyl chain at the C3 position of the β‐lactam ring. Surprisingly, the corresponding bicycle, 12d , with the PhOCH2CO side chain at C3 was inactive. Reaction models of the R39 active site gave a new insight into the geometric requirements of the conformation of potential ligands and their steric hindrance; this could help in the design of new compounds. 相似文献
The design of new amphoteric catalysts is of great interest for several industrial processes, especially those covering dehydration and dehydrogenation phenomena. Adsorption microcalorimetry was used to monitor the design of mixed oxides of zinc with Group 3 elements (aluminium, gallium, indium) with amphoteric character and enhanced specific surface area. Acid-base features were found to evolve non-linearly with the relative amounts of metal, and the strengths of the created acidic or basic sites were measured by adsorption microcalorimetry. A panel of bifunctional catalysts of various acid-base (amounts, strengths) and redox character was obtained. Besides, special interest was given to In-Zn mixed oxides for their enhanced basicity: this series of catalysts displays important basic features of high strength (q(diff) (SO? ads.) > 200 kJ mol(SO?)?1 in substantial amounts (1 - 2 μmol m(catalyst)?2), whose impact on efficiency or selectivity in catalytic dehydration/dehydrogenation can be valuable. 相似文献