Amount Of Native Protein Research Articles

Use of methods of the biocrystallomics in personification of the treatment with high diluted drugs

Background: One of most important trends in modern medicine is its transformation to personalized diagnostics and treatment. This tendency fully applies to homeopathy, especially considering that this discipline is based on the principles of individual approach to the patient. At the same time, the methods of individualization of treatment in homeopathy are relatively few. 
 Aims: Develop of methodology and methods of application biocrystalloscopic tests to personalize the homeopathic treatment. The proposed technology became previously proposed for ozone therapy practice as biocrystallomics pre-test. It includes study of the result of co-crystallization of a biological liquid of a patient with the intended dose of the drug. At the same time, the character of co-crystallization of biological liquid with several doses of tested drug or different parameters of the action of the studied factor is comparatively estimated for the purpose of individualization. Most appropriate for a particular patient believe the dose that causes optimal structuring in the dried microscopic slides obtained from a mixture of biological fluid and a solution containing a given dose of the drug. The comparison is made with a control sample of biological fluid, which has not been exposed to any effects. The optimal result is a sample that matches the control sample as much as possible. It is most preferably for biocrystallomics pre-test to use blood serum or plasma as an analyzed biological fluid.
 Methodology: To study the characteristics of the structuring of the semicroscopic slides we use the previously created system of semi-quantitative parameters. They are calculated on a straight four-point scale and include: crystallizability (semi-quantitative indicator of crystallization activity), structure index (complexity criterion for constructing crystal elements of facias), facia destruction degree (indicator of “correctness” of crystal formation) and clearity of facia marginal zone (parameter indicating the amount of native protein in the sample of biological fluid). These indicators comprehensively characterize all the main features of the process of dehydration structurization of biological substrate.
 Conclusion: It should be noted that the technique described above is universal and can be applied to any liquid or soluble compounds with potential therapeutic activity, including their ultra-high dilutions. The data obtained by us indicate the possibility of using this technology in homeopathy.

Thermally responsive silicon nanowire arrays for native/denatured-protein separation

We present our findings of the selective adsorption of native and denatured proteins onto thermally responsive, native-protein resistant poly(N-isopropylacrylamide) (PNIPAAm) decorated silicon nanowire arrays (SiNWAs). The PNIPAAm–SiNWAs surface, which shows very low levels of native-protein adsorption, favors the adsorption of denatured proteins. The amount of denatured-protein adsorption is higher at temperatures above the lower critical solution temperature (LCST) of PNIPAAm. Temperature cycling surrounding the LCST, which ensures against thermal denaturation of native proteins, further increases the amount of denatured-protein adsorption. Moreover, the PNIPAAm–SiNWAs surface is able to selectively adsorb denatured protein even from mixtures of different protein species; meanwhile, the amount of native proteins in solution is kept nearly at its original level. It is believed that these results will not only enrich current understanding of protein interactions with PNIPAAm-modified SiNWAs surfaces, but may also stimulate applications of PNIPAAm–SiNWAs surfaces for native/denatured protein separation.

Immobilization of trypsin on spent grains for whey protein hydrolysis

The aims of this work were to establish an efficient protocol for trypsin immobilization on spent grains and to assess the influence of the chosen protocol on whey protein hydrolysis. Trypsin was immobilized onto spent grain or modified spent grain, through adsorption and covalent attachment. The efficiency of immobilization and operation and storage stabilities of free and immobilized enzyme on the supports were studied. The enzyme activity, kinetic parameters and the peptide profile of the protein hydrolysates from free and immobilized enzyme were also analysed.The best activity retention was achieved with the immobilization on spent grains through multipoint covalent attachment using glycidol. This carrier showed also very good storage and operational stability (above 90%). Trypsin immobilized on spent grains showed significant activity towards whey proteins. The immobilized enzyme was slightly more stable than the free enzyme at temperatures between 50°C and 60°C allowing its use at a broader range of temperatures. The peptides formed with free enzyme and enzyme immobilized on spent grains were generally similar. However, significant differences existed in the amount of native proteins in the hydrolysates and in the relative amount of smaller peptides.

Temperature cycling stability of pre-heated acidified whey protein-stabilised o/w emulsion gels in relation to the internal surface area of the emulsion

The stability of acidified heated whey protein-based o/w emulsions against partial coalescence upon temperature cycling is investigated as a function of amount of fat, droplet size, and degree of protein denaturation in the emulsion. The data are explained in terms of a simple model that takes into account the average interdroplet distance and the amount of native protein at the interface before cycling as measures for the tendency of the emulsion to partially coalesce.

A simple model of chaperonin-mediated protein folding.

Chaperonins are oligomeric proteins that help other proteins fold. They act, according to the "Anfinsen cage" or "box of infinite dilution" model, to provide private space, protected from aggregation, where a protein can fold. Recent evidence indicates, however, that proteins are often ejected from the GroEL chaperonin in nonnative conformations, and repeated cycles of binding and ejection are needed for successful folding. Some experimental evidence suggests that GroEL chaperonins can act as folding "catalysts" in an ATP-dependent manner even when no aggregation takes place. This implies that chaperonins must somehow recognize the kinetically trapped intermediate states of a protein. A central puzzle is how a chaperonin can catalyze the folding reaction of a broad spectrum of different proteins. We propose a physical mechanism by which chaperonins can flatten the energy barriers to folding in a nonspecific way. Using a lattice model, we illustrate how a chaperonin could provide a sticky surface that helps pull apart an incorrectly folded protein so it can try again to fold. Depending on the relative sizes of the protein and the chaperonin cavity, folding can proceed both inside and outside the chaperonin. Consistent with experiments, we find that the folding rate and amount of native protein can be considerably enhanced, or sometimes reduced, depending on the amino acid sequence, the chaperonin size, and the binding and ejection rates from the chaperonin.

A simple model of chaperonin‐mediated protein folding

Chaperonins are oligomeric proteins that help other proteins fold. They act, according to the “Anfinsen cage” or “box of infinite dilution” model, to provide private space, protected from aggregation, where a protein can fold. Recent evidence indicates, however, that proteins are often ejected from the GroEL chaperonin in nonnative conformations, and repeated cycles of binding and ejection are needed for successful folding. Some experimental evidence suggests that GroEL chaperonins can act as folding “catalysts” in an ATP-dependent manner even when no aggregation takes place. This implies that chaperonins must somehow recognize the kinetically trapped intermediate states of a protein. A central puzzle is how a chaperonin can catalyze the folding reaction of a broad spectrum of different proteins. We propose a physical mechanism by which chaperonins can flatten the energy barriers to folding in a nonspecific way. Using a lattice model, we illustrate how a chaperonin could provide a sticky surface that helps pull apart an incorrectly folded protein so it can try again to fold. Depending on the relative sizes of the protein and the chaperonin cavity, folding can proceed both inside and outside the chaperonin. Consistent with experiments, we find that the folding rate and amount of native protein can be considerably enhanced, or sometimes reduced, depending on the amino acid sequence, the chaperonin size, and the binding and ejection rates from the chaperonin.

Cationization of protein antigens. I. Alteration of immunogenic properties.

We have shown that a cationized form of bovine serum albumin (BSA) produced by substituting anionic side chain carboxylic groups with aminoethylamide groups possesses unique immunologic properties. The two forms of antigen, native (nBSA) and cationized (cBSA), cross-react at the level of the B cell, as evidenced by the ability of antibody raised against one form to react with the other and by inhibition assays using ELISA. T cell cross-reactivity was also observed in proliferation assays, but the amount of cBSA required for stimulation was 500 times less than the amount of native protein needed. In vivo, cBSA produced responses which, at their optimal levels, were at least double the response to nBSA and which showed a different kinetic pattern, peaking later and lasting longer than the response to the native molecule. Moreover, antibodies were produced in response to administration of cBSA but not nBSA when given i.v. in saline, without an adjuvant. Although a mechanism for these phenomena is not yet clear, we speculate that the cBSA may have a greater affinity for antigen-presenting cells or for the T cell receptor, or that the altered structure may enhance recognition of the molecule by APC and/or helper T cells.

Studies on the Functional Properties of Food-grade Soybean Products Part I

Knowledges on chemical constituents and protein properties are fundamental prerequisites for the discussion of functional properties of food-grade soybean products. Chemical analysis was made on 43 commercial soybean products as well as on 3 gluten and 3 casein products. Principal component analysis was successfully applied for the classification of commercial products by their chemical constituents and protein properties. High correlation was observed between chemical composition and industrial processing method of soybean products. Correlation between water dispersible nitrogen and amount of native protein was unexpectedly low so that both should be taken independently as the indices for the discussion of functional properties.Flavor profiles of commercial soybean products were studied organoleptically and results were analyzed by using principal component analysis. Flavor of soybean products could be represented by two mutually independent characteristics. They are the so-called “Soybean Flavor” and “O...

Reversible dissociation of porcine ceruloplasmin into subunits

Porcine ceruloplasmin is dissociated into two subunits of equal molecular weight at 4.5 M urea in the presence of p-chloromercuribenzoate. Removal of urea by dialysis results in the formation of a homogeneous reassociated component. The component, because it is identical to the native protein in respect to molecular weight, molar extinction coefficient at 610 mμm and oxidase activity, may be the ‘native’ protein. When the samples treated with increasing concentrations of urea are dialyzed, a new component (molecular weight, 500 000) is formed in an increasing amount, and the amount of native protein is decreased. Removal of urea from the 8 M urea-treated sample results in formation of only the aggregated component. The aggregated component is devoid of oxidase activity and of blue color. To a small degree the subunits are also reassociated into the native protein upon removal of urea by gel filtration.