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1.
Some problems are considered which arise in biochemical studies on structure and function of natural oligopeptides consisting of 2–50 amino acid residues. The problems under consideration include the generation of oligopeptides from precursors, chemical structure, the role of functionally important radicals and spatial configuration, and structure-function relationships. Different types of regulation are shown mainly for oligopeptides involved in muscle contraction.Translated from Biokhimiya, Vol. 69, No. 11, 2004, pp. 1565–1573.Original Russian Text Copyright © 2004 by Zamyatnin.  相似文献   

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Oxidized plastocyanin (PC) was reduced with TyrTyrTyr and LysLysLysLysTyrTyrTyr (KKKKYYY) oligopeptides at neutral pH. The TyrTyrTyr site of the peptides provided an electron to the copper active site of PC, whereas the tetralysine site of KKKKYYY functioned as the recognition site for the negative patch of PC. The reciprocal initial rate constant (1/k(int)) increased linearly with the reciprocal TyrTyrTyr concentration and proton concentration, although the electron transfer rate decreased gradually with time. The results showed that PC was reduced by the deprotonated species of TyrTyrTyr. A linear increase of log k(int) with increase in the ionic strength was observed due to decrease in the electrostatic repulsion between negatively charged PC and deprotonated (TyrTyrTyr)(-). PC was reduced faster by an addition of KKKKYYY to the PC-TyrTyrTyr solution, although KKKKYYY could not reduce PC without TyrTyrTyr. The ESI-LCMS spectrum of the products from the reaction between PC and TyrTyrTyr showed molecular ion peaks at m/z 1015.7 and 1037.7, which suggested formation of a dimerized peptide that may be produced from the reaction of a tyrosyl radical. The results indicate that PC and the tyrosine-containing oligopeptides form an equilibrium, PC(ox)/(oligopeptide)(-)-->/<--PC(red)/(oligopeptide)(*). The equilibrium is usually shifted to the left, but could shift to the right when the produced oligopeptide radical reacts with unreacted peptides. For the reaction of PC with KKKKYYY in the absence of TyrTyrTyr, the produced KKKK(YYY)(*) radical peptide could not react with other KKKKYYY peptides, since they were positively charged. In the presence of both KKKKYYY and TyrTyrTyr, PC may interact effectively with KKKKYYY through its tetralysine site and receive an electron from its TyrTyrTyr site, where the produced KKKK(YYY)(*) may interact with TyrTyrTyr peptides.  相似文献   

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The effect of the reaction conditions (substrate concentration, enzyme dosage, and pH) on cyclodextrin production by cyclodextrin glucanotransferase from Bacillus megaterium was investigated by applying mathematical modeling methods. Adequate models were developed and they were used for determination of the optimal conditions for maximal formation of beta-cyclodextrins at minimal concentrations of a- and gamma-cydclodextrins. The main factor affecting the ratio of the products was pH of the reaction mixture. At pH 9 the enzyme formed mainly beta- and y-cyclodextrins and the ratio a:beta:gamma was 2.6:83.5:13.9; at pH 5 the ratio changed to 8.6:84.6:6.8. Mathematical models were used for determination of the conditions for maximal conversion of the substrate into cyclodextrins. 45.88% conversion of starch was achieved at 5% substrate concentration, 3.5 U/g enzyme dosage, and pH 7.4.  相似文献   

5.
Lysine oligopeptides. Preparation by ion-exchange chromatography   总被引:3,自引:0,他引:3  
The preparation of L -lysine peptides (Lysn, n = 2–14) from polyL -lysine is described. Fractionation by ion-exchange column chromatography of poly-L -lysine hydrolysates on a preparative scale resulted in 0.2–1.0 g quantities of individual members of the poly-L -lysine series. The peptides isolated proved to be analytically pure and the optical configuration was fully retained, as demonstrated by complete enzymic digestion. Peptides higher than n = 14 were also prepared. They consisted of oligolysine groups of narrow and accurately determined size distribution. Potentiometric titrations were used both to characterize the products and to demonstrate the characteristic dependence of the dissociation constants on size of the peptide.  相似文献   

6.
Chemotaxis to oligopeptides by Pseudomonas aeruginosa.   总被引:1,自引:1,他引:1       下载免费PDF全文
A number of peptides were evaluated as chemoattractants for Pseudomonas aeruginosa. Several strains recognized tri-, tetra-, penta-, and hexapeptides in a capillary tube assay. Tripeptides altered at the carboxyl terminus were good attractants, whereas tripeptides altered at the amino terminus did not serve as chemoattractants. Methionine-containing peptides were relatively poor attractants. Arginine-containing peptides gave the best responses. Reduced responses to larger peptides suggest that porin penetration is required. No extracellular peptidase activity was detected. We conclude that oligopeptides are good attractants and that specificity for chemotactic recognition of oligopeptides exists.  相似文献   

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The synthesis of the oligomeric peptides (Glu)n–Phe(NO2)–Phe (up to n = 4) by the solid-phase method is reported. Fractionation by ion-exchange column chromatography of the crude materials cleaved from the resin and subsequent amino acid analysis revealed that the desired peptides were obtained, although contaminated by several by-products whose number depends on the length of peptide chain and on the experimental conditions.  相似文献   

9.
Inhibition of superoxide by two synthetic proline-rich hexapeptides simulating the hinge region of the IgG molecule has been studied. The CPPPEL (P-Cys) peptide was active in utilizing superoxide (O2.-), while the APPPEL (P-Ala) peptide had no such activity. Spontaneous formation of clusters with six and/or eight monomers was shown for P-Cys, but not P-Ala. Preincubation of mixed P-Cys and P-Ala resulted in the appearance of a product with a high affinity for O2.-. Incubation of the mixture at 56 degrees C led to inactivation. Preincubation of P-Cys with ZnCl2 reversed the main pathway of O2.- utilization by P-Cys. The relationship of the P-Cys activity to its clusterization is discussed.  相似文献   

10.
The ciliate Tetrahymena responds very efficiently by chemoattraction to a group of trichloroacetic acid-soluble oligopeptides isolated from a commercial bioprotein from Methanococcus. When fractionated by reversed phase C18-high-pressure liquid chromatography, this group of very efficient chemoattractants turned out to consist of a heterogeneous group of oligopeptides with molecular weight ranging from 0.2 to 1.5 kDa. The peptides were very rich in the following amino acids: aspartic acid, alanine, glutamic acid, proline, glycine, lysine, and arginine. The term chemokinesis is used throughout to emphasise that chemoattraction does not necessarily include an element of orientation of cells.  相似文献   

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During screening for cyclodextrin-forming microorganisms, an alkalophilic Bacillus sp, which produced high activity of cyclodextrin glycosyltransferase, was isolated and identified as Bacillus firmus. The crude enzyme transformed starch to mainly β-and γ-cyclodextrin. The purified enzyme had an optimum pH of 7.5–8.5 and its optimum temperature was 65°C, which is the highest optimum temperature as compared to other cyclodextrin glycosyltransferases except that produced by Bacillus amyloliquefaciens. Received 06 January 1997/ Accepted in revised form 20 March 1997  相似文献   

13.
The binding of various damaged DNAs to the single-strand binding protein coded for by gene 32 from bacteriophage T4, on the one hand, and of oligopeptides containing tryptophan and lysine residues, on the other hand, is described. These molecules exhibit a higher affinity for modified DNA than for native DNA in so far as modification results in a local destabilization of the double-stranded structure of the nucleic acid. Stacking interactions between aromatic amino acids and nucleic acid bases appear to play a crucial role in the recognition of destabilized regions induced by chemical agents (carcinogens and antitumor drugs). These interactions confer to the peptide lysyl-tryptophyl-lysine an endonucleolytic activity specific for apurinic sites. From results obtained with such oligopeptides a model for the active sites of Ap-endonucleases is proposed which could account for the strategy used by the denV endonuclease from phage T4 during the first step of excision repair of pyrimidine dimers in DNA. The effect of the overall conformation of modified DNA on repair efficiency is discussed.  相似文献   

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The biological activities of two synthetic oligopeptides (His-Trp-Leu-Gln-Leu and Trp-Leu-Gln-Leu), which represent part of the primary structure of the mating hormone alpha factor from Saccharomyces cerevisiae, were studied. The peptides did not exhibit hormonal activity by themselves. However, both intensified the mating-type-specific inhibitory effect of native alpha factor on the division of haploid cells of mating type a. Random peptides or mixtures of the corresponding amino acids did not stimulate alpha factor activity. Likewise, a synthetic peptide representing another part of the alpha factor sequence was ineffective. In addition, the activity of a factor, the mating hormone produced by a cells, was not influenced by the synthetic peptides, indicating that the compounds specifically affect the interaction between alpha factor and its target cells. The analysis of the utilization of the tetrapeptide as a source of amino acids for auxotrophic a strains suggested an extracellular site of action for the observed enhancement of alpha factor activity.  相似文献   

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Ligand binding to the thrombopoietin receptor is thought to stabilize an active receptor dimer that regulates megakaryocyte differentiation and platelet formation, as well as haematopoietic stem cell renewal. By fusing a dimeric coiled coil in all seven possible orientations to the thrombopoietin receptor transmembrane (TM)-cytoplasmic domains, we show that specific biological effects and in vivo phenotypes are imparted by distinct dimeric orientations, which can be visualized by cysteine mutagenesis and crosslinking. Using functional assays and computational searches, we identify one orientation that represents the inactive dimeric state and another similar to a physiologically activated receptor. Several other dimeric orientations are identified that induce proliferation and in vivo myeloproliferative and myelodysplastic disorders, indicating the receptor can signal from several dimeric interfaces. The set of dimeric thrombopoietin receptors with different TM orientations may offer new insights into the activation of distinct signalling pathways by a single receptor and suggests that subtle differences in cytokine receptor dimerization provide a new layer of signalling regulation that is relevant for disease.  相似文献   

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By using ultrathin (350 micrometers) polyacrylamide gels, which at the end of the fractionation are pasted to filter paper and dried in an oven at 110 degrees C, and after isoelectric focusing it has been possible to detect oligopeptides in the di- to tetradecapeptide range, which could not be detected by protein staining techniques. This is achieved by developing a series of specific stains for the following amino acids: Arg, Tyr, His, Trp, Met and Cys. Except for Met and Cys, the detection limits appear to be in the order of 0.2--2 micrograms of free amino acid loaded in the gel. The Pauli reaction for His and Tyr and the Sakaguchi stain for Arg can be developed sequentially in the same gel, thus allowing the detection of four different amino acids since, under these conditions, also Trp reacts. Unfortunately, more general reactions, such as the permanganate, the 'Lowry' and the ninhydrin stains, cannot be utilized since the carrier ampholytes react very strongly with all these reagents.  相似文献   

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