Protein content,activity, and electrophoretic composition of the inhibitor-containing fraction of pea seeds

The amounts of protein and the activities of the proteinase (trypsin) inhibitors of 16 varieties of pea have been studied. The amount of protein ranged from 19.3 to 25.2% and the amidase activities of the trypsin inhibitors from 18 to 40.8 IU. It was found that the electrophoretic spectrum of the inhibitor-containing fraction of the pea seed protein consisted of 12–16 components with molecular masses of 14–89 kDa. Features of the electrophoretic spectrum of some varieties of pea have been established.V. F. Kuprevich Institute of Experimental Botany, Belorussian SSR Academy of Sciences, Minsk. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 243–247, March–April, 1989.     

Analytic calculation of phase diagrams for solutions containing colloids or globular proteins

Second-order Barker–Henderson perturbation theory gives phase diagrams for colloid and protein solutions that include stable and metastable fluid–fluid, solid–fluid, and solid–solid phases. The potential of mean force is described by a hard-sphere interacting with a Yukawa potential. Calculations for different ranges of attraction show that, as expected, fluid–fluid coexistence becomes metastable when the potential becomes short-ranged. For a very short-ranged Yukawa potential, the phase diagram shows isostructural solid–solid equilibria with a critical point. To test more simplified models, phase diagrams from second-order Barker–Henderson perturbation theory are compared with those from the random-phase approximation for the fluid phase and the van der Waals theory for the solid phase; this comparison shows significantly different phase diagrams. Moreover, with a potential of mean force with primary and secondary minima, calculations using second-order perturbation theory identify conditions where colloidal and protein solutions can present two fluid–fluid regions, each with a critical point; however, the higher-density fluid–fluid region is likely to be metastable. The analytic calculations described here may be useful for interpretation of experimental phase diagrams and for guiding design of separation processes.     

Kinetics of the qualitative and quantitative changes of various protein fractions in the ontogenesis of protoplast and hybrid cultures ofTrichoderma harzianum

The protein composition and the kinetics of the quantitative changes in the various protein fractions during the ontogenesis of protoplast and hybrid cultures ofTrichoderma harzianum have been studied. In the exponential growth phases of both cultures the highest amount of protein was contained in the water-soluble (33.0–33.2%) and salt-soluble (31.0–32.0%) fractions. The protein contents of the water-soluble fractions of the cultures studied were similar to those in meat and clover and were 3.5 times greater than in yeast.Institute of Microbiology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (3712) 41 71 29. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 209–211, March–April, 1998.     

Isolation and study of the protease-inhibitor system of wheat grain

Summary A modified method for obtaining a protease and its natural protein inhibitor from the endosperm of wheat grain is proposed. The enzyme is purified by additional precipitation of part of the ballast proteins with ammonium sulfate, and the inhibitor by two successive precipitations of the bulk of the inactive protein with the aid of TCA of different concentrations. The specific activity of the protease increases by a factor of 5–6, and the activity of the inhibitor by a factor of 3–4. Two active isoenzymes of protease have been found in each of the varieties of wheat investigated by disk eletrophoresis.Kazakh Scientific-Research Institute of Agriculture. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 642–645, September–October, 1975.     

Preparation of conjugates of peptides imitating part of the antigen-determinant section of protein VP1 of foot-and-mouth disease virus A12 with various supports

The preparation of conjugates of peptides 143–148, 153–159, 149–159, 146–159, and 143–159, imitating a section of a protein of the foot-and-mouth disease (FMD) virus of type A₁₂, with bovine serum albumin, with. a copolymer of N-vinylpyrrolidone with acrylic acid, and with a copolymer of N-vinylpyrrolidone with maleic anhydride is described. The dependence of the degree of conjugation on various factors is discussed.V. I. Lenin Tadzhik State University, Dushanbe. Institute of Immunology, Ministry of Public Health of the USSR, Moscow. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 516–519, July–August, 1990.     

Protein–protein interaction dynamics by amide H/H exchange mass spectrometry

Amide H/²H exchange detected by mass spectrometry provides a powerful tool for observing changes that occur upon protein–protein interactions. In general, it is possible to observe protection of surface amides, they become less solvent exposed when they are buried at the interface. The information thus obtained about the location of the protein–protein interface is useful for building a correct docked structure of the protein–protein complex. Examples of protein–protein interfaces that were correctly identified by such methods include the thrombin–thrombomodulin interaction and the interaction between the regulatory and catalytic subunit of protein kinase A. Amide exchange also affords a view into the subtle changes in the ensemble of states that occur upon protein modification or protein–protein binding. Examples of proteins in which amide exchange has been used to observe phosphorylation-induced changes include ERK2 and CheB. Amide exchange showed the pathway of communication between the cAMP-binding site and the catalytic subunit site within the regulatory subunit of protein kinase A. Clues as to how thrombomodulin regulates the catalytic activity of thrombin were also obtained.     

Some characteristics of the alkali-soluble protein from the red alga Furcellaria fastigiata

Summary From the red agar-bearing alga furcellaria we have isolated an alkali-soluble protein. Its aminoacid composition and N-terminal acid — arginine — have been determined. It has been shown to belong to the group of glycoproteins. One of the possible types of carbohydrate — protein bonds is a O-glycosidic bond.Odeassa Institute of Building Engineering. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 400–404, May–June, 1975.     

A lipase of the fungus Rhizopus microsporus,UzLT-1 — A glycoprotein

Summary The molecule of the lipase of the fungusRhizopus microsporus UzLT-1 consists of protein and nonprotein moieties. The nonprotein moiety is formed by carbohydrates.The carbohydrate moiety is readily separated from the protein moiety, which suggested a noncovalent bond of the sugars with the protein fraction.Department of Microbiology of the Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 265–267, March–April, 1977.     

Presynaptic and postsynaptic neurotoxins. Investigation of the structures of the immune recognition sections

This paper discusses literature reports over the last 12 years and the results of our own experimental investigations of the structures of the immune recognition regions of protein antigens — in particular presynaptic and postsynaptic neurotoxins from spider and snake venoms.Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 22–40, January–February, 1998.     

The precipitation of isolated cotton-plant protein

Summary The optimum condition for the precipitation of the protein from an extract obtained by treating cottonseed meal with 5% NH₄Cl is its precipitation at room temperature with 10% HCl at pH 3–3.5.Institute of the Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 116–118, January–February, 1978.     

Some physicochemical parameters of transferrin isolated from rat blood serum

Chromatography on ion-exchange resins has yielded a homogeneous iron-containing glycoprotein — transferrin. The amino acid composition of the protein has been shown, it has been established that the N-terminal amino acid of the protein is threonine, and the sedimentation coefficient has been calculated, being 5.8 S.Institute of Bioorganic Chemistry, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 89–92, January–February, 1980.     

On the hypothetical protein F154 of the TTV1 virus/Thermoproteus Tenax. Part II: Synthesis of the trieicosapeptide,corresponding to the protein sequence 79–101

Summary For the identification of a protein predicted by DNA sequence analysis of the TTV1 virus from the archaebacteriumThermoproteus tenax, the trieicosapeptide H-Thr-Pro-Thr-Pro-Thr-Pro-Thr-Tyr-Asp-Ile-Thr-Tyr-Val-Val-Phe-Asp-Val-Thr-Pro-Ser-Pro-Thr-Pro-OH, corresponding to the protein fragment 79–101, was prepared by conventional methods of peptide synthesis. This sequence portion may possibly represent a suitable protein specific immunepitope.

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Zur Hypothese eines TTV1 Virus/Thermoproteus tenax F154-Proteins. Teil II: Synthese des Protein-fragments 79–101

Zusammenfassung Für den Nachweis der Expression des Proteins F154 — nach einer Sequenzanalyse des Genoms des TTV1 Virus im ArchaebakteriumThermoproteus tenax postuliert — wurde das Peptid H-Thr-Pro-Thr-Pro-Thr-Pro-Thr-Tyr-Asp-Ile-Thr-Tyr-Val-Val-Phe-Asp-Val-Thr-Pro-Ser-Pro-Thr-Pro-OH (Proteinfragment 79–101) mit Hilfe konventioneller Peptidsynthese hergestellt. Diese Peptidsequenz sollte ein geeignetes proteinspezifisches Immunepitop darstellen.

     

Purification of nerve growth factor from the venom of the Central Asian cobraNaja oxiana

A highly purified electrophoreticaly homogeneous protein with a NGF activity of 10·10⁵ BU/mg of protein have been isolated from the venom of the Central Asian cobra by gel-filtration and ion-exchange chromatography followed by preparative isolectric focusing in a thin layer of Sephadex. It has been shown that the NGF isolated is characterized by a molecular weight in the range of 20–30 kD and a pI value of about 7.0.Institute of Biochemistry, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii. No. 4, pp. 546–551, July–August, 1985.     

Genetically engineered peptide fusions for improved protein partitioning in aqueous two-phase systems: Effect of fusion localization on endoglucanase I of Trichoderma reesei

Genetic engineering has been used for fusion of the peptide tag, Trp–Pro–Trp–Pro, on a protein to study the effect on partitioning in aqueous two-phase systems. As target protein for the fusions the cellulase, endoglucanase I (endo-1,4-β- -glucan-4-glucanohydrolase, EC 3.2.1.4, EGI, Cel7B) of Trichoderma reesei was used. For the first time a glycosylated two-domain enzyme has been utilized for addition of peptide tags to change partitioning in aqueous two-phase systems. The aim was to find an optimal fusion localization for EGI. The peptide was (1) attached to the C-terminus end of the cellulose binding domain (CBD), (2) inserted in the glycosylated linker region, (3) added after a truncated form of EGI lacking the CBD and a small part of the linker. The different constructs were expressed in the filamentous fungus T. reesei under the gpdA promoter from Aspergillus nidulans. The expression levels were between 60 and 100 mg/l. The partitioning behavior of the fusion proteins was studied in an aqueous two-phase model system composed of the thermoseparating ethylene oxide (EO)–propylene oxide (PO) random copolymer EO–PO (50:50) (EO₅₀PO₅₀) and dextran. The Trp–Pro–Trp–Pro tag was found to direct the fusion protein to the top EO₅₀PO₅₀ phase. The partition coefficient of a fusion protein can be predicted with an empirical correlation based on independent contributions from partitioning of unmodified protein and peptide tag in this model system. The fusion position at the end of the CBD, with the spacer Pro–Gly, was shown to be optimal with respect to partitioning and tag efficiency factor (TEF) was 0.87, where a fully exposed tag would have a TEF of 1.0. Hence, this position can further be utilized for fusion with longer tags. For the other constructs the TEF was only 0.43 and 0.10, for the tag fused to the truncated EGI and in the linker region of the full length EGI, respectively.     

Some functional properties of cotton protein hydrolysates

A comparative study of the solubility and emulsifying properties of cotton protein hydrolysates produced by the enzyme preparation Pektofoetidin has shown that the optimum functional properties are possessed by hydrolysates with a degree of proteolysis of 8–13%.Institute of Chemistry of Plant Substances, Academy of Sciences of the Uzbek SSR, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 520–524, July–August, 1990.     

Modification of the reserve proteins during the germination of cotton seeds

The hydrolysis of the 11S globulin begins in the early stages of the germination of cotton seeds and is complete during the first 4–5 days. During the first 3 days, only limited hydrolysis of the reserve protein takes place, and only on the 4th day does it undergo far-reaching hydrolysis, after which the protein cannot be detected either by immunochemical methods or by electrophoresis and analytical ultracentrifugation.Institute of Chemistry of Plant Substances, Academy of Sciences of the Uzbekistan Republic, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 443–445, 447, May–June, 1993.     

Protein kinase from cotton seeds

A Ca²⁺-dependent protein kinase C, active at pH 6.5–8.0 has been found in cotton seeds for the first time. The localization of the enzyme in the seeds has been established and some of its properties are described (stability in various media, capacity for performing the phosphorylation of various substrates, activation by calcium ions). Highly active preparations of cottonseed protein kinase C have been isolated by biospecific chromatography.Institute of Bioorganic Chemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 255–259, March–April, 1989.     

Inhomogeneity in Distribution and Conformation of Bovine Serum Albumin in Sol–Gel: A Closer Look with a Near-Infrared Multispectral Imaging Technique

Concentration distributions and conformations of bovine serum albumin (BSA) entrapped in sol–gels were successfully determined for the first time by use of the recently developed NIR multispectral imaging instrument. It was found that BSA molecules were inhomogeneously distributed within the sol–gel matrix, independent of its concentration. At relatively high concentration (366 mg/mL) the encapsulation process does not seem to produce any observable changes in the conformation of BSA. However, when the concentration of BSA was decreased to 220 mg/mL, pronounced changes in the spectra of the protein were observed as a function of (sol–gel reaction) time. The observed inhomogenity might be traced to both a non-uniform concentration distribution of BSA and changes in protein native conformation resulting from interactions between the charged protein and the silicate anions. Results obtained seem to suggest that the protein molecules might adapt themselves to accommodate evolving geometry and sites of the silica network.     

Water-soluble ethylene-binding proteins of the cotton plant: Isolation and functional activity

A protein possessing a high affinity for ethylene has been isolated from cottonplant shoots by hydrophobic chromatography on Polikhrom-1 and gel chromatography on TSK gel HW-50F. A number of proofs of the receptor nature of the protein isolated are given.S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax 627071. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 92–96, January–February, 1994.     

Biochemical and immunochemical study of a protein preparation from the culture liquid ofYersinia pseudotuberculosis

Using ultrafiltration methods of isolation, a culture liquid ofY. pseudotuberculosis has yielded a protein fraction with a molecular mass of not less than 80–100 kDa which possesses hemagglutinating activity and the properties of an antireceptor. It has been shown for the first time by the methods of affinity chromatography, indirect hemagglutination, and indirect IEA that this protein preparation from aY. pseudotuberculosis culture interacts specifically with the Fab₂ fragment of normal human IgG.Pacific Ocean Institute of Bioorganic Chemistry, Far Eastern Division of the Russian Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 784–787, November–December, 1994.