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Monomeric cyt c has been reported to bind to the mitochondrial membrane by electrostatic and hydrophobic interactions with anionic phospholipids. We have previously shown that domain‐swapped oligomeric cyt c retains the secondary structure of the monomer, and its surface possesses a larger area and more charges compared to the monomer. However, the effect of oligomerization of cyt c on cells has yet to be revealed. Herein, we investigated the interaction of oligomeric cyt c with anionic phospholipid‐containing vesicles and the outer membrane of HeLa cells. Oligomeric cyt c interacted more strongly than monomeric cyt c with anionic phospholipid‐containing vesicles and the outer membrane of HeLa cells. Oligomeric cyt c induced lateral phase separation of lipids in LUVs and GUVs, thereby leading to membrane disruption, whereas monomeric cyt c did not. Morphological changes in HeLa cells resulted from interaction with oligomeric cyt c, but little from interaction with the monomer. These results show that domain‐swapped oligomeric proteins might exhibit properties different to those of monomer in cell systems.  相似文献   

3.
Three out of four subtypes of arrestin proteins expressed in mammals self-associate, each forming oligomers of a distinct kind. Monomers and oligomers have different subcellular localization and distinct biological functions. Here we summarize existing evidence regarding arrestin oligomerization and discuss specific functions of monomeric and oligomeric forms, although too few of the latter are known. The data on arrestins highlight biological importance of oligomerization of signaling proteins. Distinct modes of oligomerization might be an important contributing factor to the functional differences among highly homologous members of the arrestin protein family.  相似文献   

4.
The tetraalcohol 2,3,5,6-endo,endo,endo,endo-tetrakis(hydroxymethyl)bicyclo[2.2.1]heptane (tetol, 1) has been prepared and crystallises readily as the lithium(I) complex [Li(1)2]Cl, forming an oligomeric multi-chain structure in which pairs of alcohols from two crystallographically independent tetol molecules bind lithium ions tetrahedrally. However, formation of monomeric structures in solution is inferred from electrospray mass spectroscopy, which has also shown evidence of exchange of lithium ion in the complexed species by added alkaline earth ions.  相似文献   

5.
The Application of Gel Permeation Chromatography to the Investigation of Thermal-Oxidatively Treated Oils Principle and apparatus used in gel permeation chromatography (GPC) are given along with methods for the determination of dimeric, trimeric and higher oligomeric triglycerides in frying fats that are subjected to extreme thermal oxidation. Separation of a frying fat into monomeric and oligomeric triglycerides by means of preparative GPC followed by analyses of their products of saponification revealed that dimeric fatty acids occur only in the fraction of oligomeric triglycerides. Accordingly, the dimerization reaction in the presence of atmospheric oxygen consists entirely of an intermolecular reaction. Under various conditions (at 240°C in the presence of air, at 240°C under argon, and under partial exposure to both light and air) all the oligomerization reactions obey the reaction law of the first order, although their rates are different. In the investigation of animal fats by GPC tallow and butter were found to contain none, whereas the fat in cheese, chicken and lard were found to contain upto 1% dimeric triglycerides.  相似文献   

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7.
Antenna proteins play a major role in the regulation of light-harvesting in photosynthesis. However, less is known about a possible link between their sizes (oligomerization state) and fluorescence intensity (number of photons emitted). Here, we used a microscopy-based method, Fluorescence Correlation Spectroscopy (FCS), to analyze different antenna proteins at the particle level. The direct comparison indicated that Chromera Light Harvesting (CLH) antenna particles (isolated from Chromera velia) behaved as the monomeric Light Harvesting Complex II (LHCII) (from higher plants), in terms of their radius (based on the diffusion time) and fluorescence yields. FCS data thus indicated a monomeric oligomerization state of algal CLH antenna (at our experimental conditions) that was later confirmed also by biochemical experiments. Additionally, our data provide a proof of concept that the FCS method is well suited to measure proteins sizes (oligomerization state) and fluorescence intensities (photon counts) of antenna proteins per single particle (monomers and oligomers). We proved that antenna monomers (CLH and LHCIIm) are more “quenched” than the corresponding trimers. The FCS measurement thus represents a useful experimental approach that allows studying the role of antenna oligomerization in the mechanism of photoprotection.  相似文献   

8.
对比分析了3批聚合度为1000的聚氯乙烯(PVC)树脂的热稳定性、相对分子质量及其分布、偏二氯乙烯链节含量、分子链双键的相对含量及重金属离子含量.结果发现,热稳定性差的树脂双键相对含量较高,重金属离子相对含量明显高出正常批次料;同时还检测出了偏二氯乙烯链节.研究认为,引起配混料和医用粒料颜色发暗、雾度升高、制品黄色指数高的主要原因首先是树脂重金属离子含量高,对树脂老化起到了催化作用;其次是原料单体中的杂质参与了聚合,形成了分子链缺陷,导致热稳定性下降.  相似文献   

9.
Enzymes catalyzing the hydrolysis of the N-glycosidic bond in nucleosides and other ribosides (N-ribohydrolases, NHs) with diverse substrate specificities are found in all kingdoms of life. While the overall NH fold is highly conserved, limited substitutions and insertions can account for differences in substrate selection, catalytic efficiency, and distinct structural features. The NH structural module is also employed in monomeric proteins devoid of enzymatic activity with different physiological roles. The homo-oligomeric quaternary structure of active NHs parallels the different catalytic strategies used by each isozyme, while providing a buttressing effect to maintain the active site geometry and allow the conformational changes required for catalysis. The unique features of the NH catalytic strategy and structure make these proteins attractive targets for diverse therapeutic goals in different diseases.  相似文献   

10.
A correlation of the catalytic activity for anodic chlorine evolution of platinum group metals to the nature of the surface film formed during chlorine evolution in a sodium chloride solution was studied by X-ray photoelectron spectroscopy. The change in the surface film with increasing potential was found on platinum, including an increase in the cationic valence. This seemed responsible for the decrease in the activity for chlorine evolution on platinum in the high potential region. Increasing potential did not result in the appreciable increase in the cationic valence in the surface film on the other platinum group metals. Replacement of hydroxyl ions in the surface film by chloride ions became easier in the order of rhodium, iridium and palladium, and the activity for anodic chlorine evolution increased in this order due possibly to an increase in the amount of chloride ions in the film which seemed to be one of the reactants in the rate determining electrochemical desorption of adsorbed chlorine atom. Chlorine molecules adsorbed on the surface film were also found. It was assumed that the activity for anodic chlorine evolution might be low when the metal surface was covered by a large amount of molecular chlorine which was the reaction product.  相似文献   

11.
A mutation at the dimer interface of Plasmodium falciparum triosephosphateisomerase (PfTIM) was created by mutating a tyrosine residueat position 74, at the subunit interface, to glycine. Tyr74is a critical residue, forming a part of an aromatic clusterat the interface. The resultant mutant, Y74G, was found to haveconsiderably reduced stability compared with the wild-type protein(TIMWT). The mutant was found to be much less stable to denaturingagents such as urea and guanidinium chloride. Fluorescence andcircular dichroism studies revealed that the Y74G mutant andTIMWT have similar spectroscopic properties, suggestive of similarfolded structures. Further, the Y74G mutant also exhibited aconcentration-dependent loss of enzymatic activity over therange 0.1–10 µM. In contrast, the wild-type enzymedid not show a concentration dependence of activity in thisrange. Fluorescence quenching of intrinsic tryptophan emissionwas much more efficient in case of Y74G than TIMWT, suggestiveof greater exposure of Trp11, which lies adjacent to the dimerinterface. Analytical gel filtration studies revealed that inY74G, monomeric and dimeric species are in dynamic equilibrium,with the former predominating at low protein concentration.Spectroscopic studies established that the monomeric form ofthe mutant is largely folded. Low concentrations of urea alsodrive the equilibrium towards the monomeric form. These studiessuggest that the replacement of tyrosine with a small residueat the interface of triosephosphate isomerase weakens the subunit–subunitinteractions, giving rise to structured, but enzymatically inactive,monomers at low protein concentration.  相似文献   

12.
Aptameric sensors can bind molecular targets and produce output signals, a phenomenon that is used in bioassays. In some cases, it is important to distinguish between monomeric and oligomeric forms of a target. Here, we propose a strategy to convert a monomer/oligomer‐nonselective sensor into an oligomer‐selective sensor. We designed an aptazyme that produced a high fluorescent output in the presence of oligomeric α‐synuclein (a molecular marker of Parkinson's disease) but not its monomeric form. The strategy is potentially useful in the design of point‐of‐care tests for the diagnosis of neurodegenerative diseases.  相似文献   

13.
Proper protein folding is a prerequisite for protein stability and enzymatic activity. Although directed evolution can be a powerful tool to investigate enzymatic function and to isolate novel activities, well‐designed libraries of folded proteins are essential. In vitro selection methods are particularly capable of searching for enzymatic activities in libraries of trillions of protein variants, yet high‐quality libraries of well‐folded enzymes with such high diversity are lacking. We describe the construction and detailed characterization of a folding‐enriched protein library based on the ubiquitous (β/α)8 barrel fold, which is found in five of the six enzyme classes. We introduced seven randomized loops on the catalytic face of the monomeric, thermostable (β/α)8 barrel of glycerophosphodiester phosphodiesterase (GDPD) from Thermotoga maritima. We employed in vitro folding selection based on protease digestion to enrich intermediate libraries containing three to four randomized loops for folded variants, and then combined them to assemble the final library (1014 DNA sequences). The resulting library was analyzed by using the in vitro protease assay and an in vivo GFP‐folding assay; it contains ~1012 soluble monomeric protein variants. We isolated six library members and demonstrated that these proteins are soluble, monomeric and show (β/α)8‐barrel fold‐like secondary and tertiary structure. The quality of the folding‐enriched library improved up to 50‐fold compared to a control library that was assembled without the folding selection. To the best of our knowledge, this work is the first example of combining the ultra‐high throughput mRNA display method with selection for folding. The resulting (β/α)8 barrel libraries provide a valuable starting point to study the unique catalytic capabilities of the (β/α)8 fold, and to isolate novel enzymes.  相似文献   

14.
An electrochemical study of prestressed steel in saturated Ca(OH)2 aqueous solutions (pH 12) was carried out in the absence and in the presence of chloride ions, in such a concentration that simulates the composition of seawater. Cyclic voltammetry, linear sweep voltammetry, open circuit potential transients, atomic absorption spectroscopy and scanning electron microscopy coupled to electron diffraction spectroscopy were employed. The linear polarisation curves analysis led to the determination of polarisation resistance, R p, corrosion potential, E corr, corrosion current density, j corr , Tafel slopes, breakdown potential, E b and repassivation potential E repass. A linear dependence of the breakdown potential, E b, on the square root of scan rate was obtained, according to the Point Defect Model (PDM). A crossover characteristic of the nucleation processes was observed in the presence of chloride ions. SEM/EDS studies revealed, as expected, a strong influence of the presence of chloride ions observed in the transpassive and the active regions. In conclusion, chloride ions contribute to enhance the corrosion of steel, most probably due to their adsorption on both the active and the passive electrode surfaces.  相似文献   

15.
Hexagonal mesoporous silica (HMS) catalysts post-synthetically doped with vanadia oxo-species were characterized by means of XRD, UV-Vis spectroscopy, H2-TPR and studied in oxidative dehydrogenation of propane (ODH). The relationship between catalytic activity in ODH and the presence of different vanadia-oxo species (monomeric, oligomeric and oxide-like species) was suggested. Monomeric VOx species are responsible for high catalytic activity and selectivity, oligomeric species containing V-O-V bond are active but non-selective to propene and oxide-like VOx particles are significantly less active and selective.  相似文献   

16.
Individual dimer, trimer, and tetramer constituents that comprise the higher molecular weight (mol wt), mesophase-forming fraction of M-50 petroleum pitch were characterized in terms of polycyclic aromatic hydrocarbon (PAH) backbone structure, the extent of alkylation, and the nature of the bonds connecting the monomeric “building blocks” comprising the oligomeric species. Isolation of individual oligomeric constituents for subsequent analysis and identification was made possible by applying our two-step, sequential fractionation technique of dense-gas/supercritical extraction (DGE) followed by prep-scale gel permeation chromatography (GPC). MALDI and MALDI-post source decay (PSD) mass spectrometry of the narrow mol wt fractions obtained indicate that the major oligomeric constituents are grouped in terms of well-defined molecular weight distributions, with each composed of base monomer PAH backbone structure(s) (e.g., pyrene, triphenylene, benzopyrenes, chrysene, and benz[a]anthracene) possessing various degrees (typically 1–4/monomer unit) of methylation. UV–Vis analysis indicates that the oligomerization process occurs via a condensation reaction, with the loss of four hydrogens as a nonalternant, 5-membered PAH ring forms to join the reacting monomer units. FT-IR spectroscopy indicates that this reaction results in relatively uncondensed structures over the entire range of oligomers investigated. Analogous results to those given above were obtained for the thermal polymerization of an anthracene pitch.  相似文献   

17.
Caseinolytic protease P (ClpP) is a tetradecameric peptidase that assembles with chaperones such as ClpX to gain proteolytic activity. Acyldepsipeptides (ADEPs) are small-molecule mimics of ClpX that bind into hydrophobic pockets on the apical site of the complex, thereby activating ClpP. Detection of ClpP has so far been facilitated with active-site-directed probes which depend on the activity and oligomeric state of the complex. To expand the scope of ClpP labeling, we took a stepwise synthetic approach toward customized ADEP photoprobes. Structure–activity relationship studies with small fragments and ADEP derivatives paired with modeling studies revealed the design principles for suitable probe molecules. The derivatives were tested for activation of ClpP and subsequently applied in labeling studies of the wild-type peptidase as well as enzymes bearing mutations at the active site and an oligomerization sensor. Satisfyingly, the ADEP photoprobes provided a labeling readout of ClpP independent of its activity and oligomeric state.  相似文献   

18.
Cyclic oligomeric carbonates are prepared by a hydrolysis/condensation reaction from aromatic bis(chloroformate)s. Three methods for convenient preparation of bis(chloroformate)s have been developed: (1) use of diethylaniline to scavenge HCl, in a modification of an earlier procedure; (2) low pH, low temperature interfacial condensation of bisphenols with phosgene; and (3) use of Ca(OH)2 in interfacial condensation with phosgene. Reaction parameters which control formation of monomeric bis(chloroformate)s versus higher oligomerization include temperature, pH, and rate of phosgene addition. For water-soluble bisphenols such as hydroquinone, the phase ratio of water to CH2Cl2 can also be important.  相似文献   

19.
Riboflavin serves as a precursor for flavocoenzymes (FMN and FAD) and is essential for all living organisms. The two committed enzymatic steps of riboflavin biosynthesis are performed in plants by bifunctional RIBA enzymes comprised of GTP cyclohydrolase II (GCHII) and 3,4-dihydroxy-2-butanone-4-phosphate synthase (DHBPS). Angiosperms share a small RIBA gene family consisting of three members. A reduction of AtRIBA1 expression in the Arabidopsis rfd1mutant and in RIBA1 antisense lines is not complemented by the simultaneously expressed isoforms AtRIBA2 and AtRIBA3. The intensity of the bleaching leaf phenotype of RIBA1 deficient plants correlates with the inactivation of AtRIBA1 expression, while no significant effects on the mRNA abundance of AtRIBA2 and AtRIBA3 were observed. We examined reasons why both isoforms fail to sufficiently compensate for a lack of RIBA1 expression. All three RIBA isoforms are shown to be translocated into chloroplasts as GFP fusion proteins. Interestingly, both AtRIBA2 and AtRIBA3 have amino acid exchanges in conserved peptides domains that have been found to be essential for the two enzymatic functions. In vitro activity assays of GCHII and DHBPS with all of the three purified recombinant AtRIBA proteins and complementation of E. coli ribA and ribB mutants lacking DHBPS and GCHII expression, respectively, confirmed the loss of bifunctionality for AtRIBA2 and AtRIBA3. Phylogenetic analyses imply that the monofunctional, bipartite RIBA3 proteins, which have lost DHBPS activity, evolved early in tracheophyte evolution.  相似文献   

20.
In this study, chemical recycling products of waste poly(ethylene terephthalate) with oligoesters were used as new plasticizers for poly(vinyl chloride) (PVC). The preparation conditions of the dry blend mixtures of the suspension PVC containing synthesised plasticizers were similar to the conditions of the preparing mixtures with commercial plasticizers. The plasticization efficiency of PVC plasticizers was then examined by analysis of the mechanical, physical and chemical properties, as well as the thermal resistance and migration of plasticizer molecules from polymer matrix. Test results proved that compositions with synthesised oligomeric plasticizers possessed similar or better properties than those containing commercial oligomeric plasticizers and much better properties than those having monomeric plasticizers. Thermal stabilities of the proposed plasticizers were higher than those of the commercial plasticizers either monomeric (bis(2-ethylhexyl)phthalate) or oligomeric, despite the fact that the synthesised oligoesters did not contain any antioxidant. The best properties, especially low volatility, very good mechanical properties, low migration were resulted of the transesterification of the waste PET with oligoesters based on adipic acid, triethylene glycol and 2-ethylhexanol which were selected as plasticizers synthesised on the technical scale. The tested plasticized PVC compositions possessed very good tear resistance, tensile strength, decrease of weight loss after 168 h at 80 °C and low migration. Processing properties of PVC compositions containing these synthesised plasticizers confirmed their effectiveness in these compositions for extrusion process.  相似文献   

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