Spatial structure of myelopeptides: II. Conformational analysis of MP-4, MP-5, and MP-6

Theoretical conformational analysis was used to study the spatial structure and conformational properties of myelopeptides, bone-marrow peptide mediators. The low-energy conformations of myelopeptides MP-4 (Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro), MP-5 (Val-Val-Tyr-Pro-Asp), and MP-6 (Val-Asp-Pro-Pro) were found; the values of dihedral angles of backbone and side chains of the amino acid residues were determined; and the energies of intra- and interresidual interactions were estimated.__________Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 2, 2005, pp. 140–146.Original Russian Text Copyright © 2005 by Ismailova, Akhmedov, Abbasli, Godjaev.     

Spatial structure of myelopeptides: I. conformational analysis of MP-1, MP-2, and MP-3

Theoretical conformational analysis was used to study the spatial structure and conformational properties of myelopeptides, bone-marrow peptide mediators. The low-energy conformations of three hexapeptides MP-1 (Phe-Leu-Gly-Phe-Pro-Thr), MP-2 (Leu-Val-Val-Tyr-Pro-Trp), and MP-3 (Leu-Val-Cys-Tyr-Pro-Gln) were found, the values of dihedral angles of the backbone and side chains of the amino acid residues con-stituting these peptides were determined, and the energies of intra- and interresidual interactions were estimated.Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 1, 2005, pp. 31–38.Original Russian Text Copyright © 2005 by Akhmedov, Ismailova, Abbasli, Akhmedov, Godjaev.     

Spatial structure of myelopeptides: I. Conformational analysis of MP-1, MP-2, and MP-3

Theoretical conformational analysis was used to study the spatial structure and conformational properties of myelopeptides, bone-marrow peptide mediators. The low-energy conformations of three hexapeptides MP-1 (Phe-Leu-Gly-Phe-Pro-Thr), MP-2 (Leu-Val-Val-Tyr-Pro-Trp), and MP-3 (Leu-Val-Cys-Tyr-Pro-Gln) were found, the values of dihedral angles of the backbone and side chains of the amino acid residues constituting these peptides were determined, and the energies of intra- and interresidual interactions were estimated.     

Effect of the mineral-ammonia preparations MP-15 and MP-7 on nitrogen metabolism in cows

It has been studied how MP-15 and MP-7 preparations fed on to lactating cows affect the indices of acid-base equilibrium of their blood, nitrogen metabolism, milk yield and milk quality. It is shown that feeding on MP-15 and MP-7 preparations to cows of test groups has no negative effect on the acid-base equilibrium indices, decreases the ammonia concentration by 38.5 and 54.9%, respectively, increases the glutamine content in blood by 23.5 and 29.4%, respectively, does not affect the pyruvate, glutamate, oxaloacetate concentration and alanine-transaminase activity, increases the average daily milk yields by 2.1 and 2.3 l, respectively, and does not deteriorate the composition and physicochemical indices of milk quality.     

Regulation of the peritoneal macrophage functional activity by the MP-5 and MP-6 myelopeptides under stress

In mice, two-hour immobilization stress inhibited zymosan-induced production by macrophages of the oxygen radicals and cytokine IL-1β. After myelopeptides MP-5 and MP-6 were administered into mice, the stress-induced inhibition of the reactive oxygen species (ROS) and IL-1β was abrogated. MP-5 peptide stimulated spontaneous ROS production by macrophages and reduced IL-10 production under stress. Thus, under in vivo conditions and under stress, the effect of MP-5 and MP-6 myelopeptides modulates the peritoneal macrophage activity.     

Genetic Subpopulations of Rift Valley Fever Virus Strains ZH548 and MP-12 and Recombinant MP-12 Strains

Rift Valley fever virus strain MP-12 was generated by serial plaque passages of parental strain ZH548 12 times in MRC-5 cells in the presence of a chemical mutagen, 5-fluorouracil. As a result, MP-12 encoded 4, 9, and 10 mutations in the S, M, and L segments, respectively. Among them, mutations in the M and L segments were responsible for attenuation, while the MP-12 S segment still encoded a virulent phenotype. We performed high-throughput sequencing of MP-12 vaccine, ZH548, and recombinant MP-12 (rMP-12) viruses. We found that rMP-12 contains very low numbers of viral subpopulations, while MP-12 and ZH548 contain 2 to 4 times more viral genetic subpopulations than rMP-12. MP-12 genetic subpopulations did not encode the ZH548 sequence at the 23 MP-12 consensus mutations. On the other hand, 4 and 2 mutations in M and L segments of MP-12 were found in ZH548 subpopulations. Thus, those 6 mutations were no longer MP-12-specific mutations. ZH548 encoded several unique mutations compared to other Egyptian strains, i.e., strains ZH501, ZH1776, and ZS6365. ZH548 subpopulations shared nucleotides at the mutation site common with those in the Egyptian strains, while MP-12 subpopulations did not share those nucleotides. Thus, MP-12 retains unique genetic subpopulations and has no evidence of reversion to the ZH548 sequence in the subpopulations. This study provides the first information regarding the genetic subpopulations of RVFV and shows the genetic stability of the MP-12 vaccine manufactured in MRC-5 cells.     

Synthesis and antitumor properties of the myelopeptide MP-1

The bone marrow-derived peptide Phe-Leu-Gly-Phe-Pro-Thr (MP-1) has been synthesized by the classical methods of peptide chemistry in solution, and its antitumor properties have been studied. It has been shown that MP-1 enhances the efficacy of the cytostatic therapy of lympholeukosis P388, increases the latent period of the growth of P388 tumors implanted in irradiated mice, and reduces the recurrence of the breast adenocarcinoma Ca-755 in mice after the surgery.     

Myelopeptide MP-5 and fluorescent derivatives: Synthesis and biological activity

The Val-Val-Tyr-Pro-Asp bone marrow peptide (MP-5) and its analogue (MP-5-Lys) were synthesized. Fluorescent derivatives, Ftc-MP-5 and MP-5-Lys(Ftc), were prepared. The iological activity of MP-5 and MP-5-Lys was studied in vitro and in vivo. The MP-5 peptide caused 60–84% inhibition of growth of the following mouse cancers: lymphatic leukemia P388, melanoma B-16, and cervical carcinoma CUC-5. These peptides also restored functional activity of T-lymphocytes that was inhibited by metabolic products of the HL-60 leukemic cell line. MP-5-Lys(Ftc) was shown to preserve the functional properties of MP-5 towards T-lymphocytes, but Ftc-MP-5 was practically inactive.     

Synthesis and properties of the retro-analogue of myelopeptide MP-2

The bone marrow myelopeptide MP-2 (Leu-Val-Val-Tyr-Pro-Trp), exhibiting antitumor activity, and its retro-analogue (Trp-Pro-Tyr-Val-Val-Leu) were synthesized, and their properties were studied. The in vitro and in vivo activities of retro-MP-2 were comparable with those of MP-2. Both peptides equally restored the functional activity of T-lymphocytes inhibited by toxins released by HL-60 cells and inhibited by 70-82% the growth of various types of transplantable solid tumors: Ca-755 adenocarcinoma of the mammary gland, Lewis adenocarcinoma of the lung, and S180 sarcoma. The positions and intensities of the Cotton effects in CD spectra of the MP-2 peptide and its retro-analogue in various solvents are almost indistinguishable. The positions of extrema and integral intensities of the amide I and amide A bands in IR spectra of both peptides were practically identical. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.     

Purification and characterization of mutanase produced by Paenibacillus curdlanolyticus MP-1

Mutanases are enzymes that catalyze hydrolysis of α-1,3-glucosidic bonds in various α-glucans. One of such glucans, mutan, which is synthesized by cariogenic streptococci, is a major virulence factor for induction of dental caries. This means that mutan-degrading enzymes have potential in caries prophylaxis. In this study, we report the purification, characterization, and partial amino acid sequence of extracellular mutanase produced by the MP-1 strain of Paenibacillus curdlanolyticus, bacterium isolated from soil. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme showed a single protein band of molecular mass 134 kD, while native gel filtration chromatography confirmed that the enzyme was a monomer of 142 kD. Mutanase showed a pH optimum in the range from pH 5.5 to 6.5 and a temperature optimum around 40-45°C. It was thermostable up to 45°C, and retained 50% activity after 1 hr at 50°C. The enzyme was fully stable at a pH range of 4 to 10. The enzyme activity was stimulated by the addition of Tween 20, Tween 80, and Ca2?, but it was significantly inhibited by Hg2?, Ag?, and Fe2?, and also by p-chloromercuribenzoate, iodoacetamide, and ethylenediamine tetraacetic acid (EDTA). Mutanase preparation preferentially catalyzed the hydrolysis of various streptococcal mutans and fungal α-1,3-glucans. It also showed binding activity to insoluble α-1,3-glucans. The N-terminal amino acid sequence was NH?-Ala-Gly-Gly-Thr-Asn-Leu-Ala-Leu-Gly-Lys-Asn-Val-Thr-Ala-Ser-Gly-Gln. This sequence indicated an analogy of the enzyme to α-1,3-glucanases from other Paenibacillus and Bacillus species.     

灵芝MP-01菌株液体发酵培养基的筛选

对灵芝MP-01菌株在三种培养基上的生长性状进行了研究,并对其液体发酵过程中的pH值变化、发酵全液的多糖含量变化、菌丝生物量及发酵培养性状等进行了观测。结果表明,玉米面培养基是灵芝MP-01菌株液体发酵的适宜培养基,其适宜的终止发酵时间在培养的第6 d,此时发酵液多糖含量达到10.32 mg/mL。     

Studies on the biosynthesis of bialaphos (SF-1293) Part 3. Production of phosphinic acid derivatives,MP-103, MP-104 and MP-105, by a blocked mutant of Streptomyceshygroscopicus SF-1293 and their roles in the biosynthesis of bialaphos

During biosynthetic studies on bialaphos to reveal the formation mechanisms of carbon-phosphorous bonds in detail, three new metabolites containing a HPC bond structure were isolated from the fermentation broth of a mutant of $\text{Streptomyces}\text{hygroscopicus}$ SF-1293. Based on the spectroscopic analysis, the structures of these compounds have been determined as shown in Fig. 1. Transformation experiments of these metabolites to bialaphos suggested that the reduction of the phosphorous atom in phosphate will take place at an early biosynthetic stage.     

Effect of preparation MP-15 on the productivity and metabolism of lactating cows

Activation possibilities of redox and biosynthetic processes are studied for lactic cows with their ration supplemented by mineral-ammonium preparation MII-15. In the blood of cows from the experimental group the amount of a number of glycolysis, biocarboxylic cycle, ammonium nitrogen exchange products as well as activity of overamination enzymes vary as compared to control, which testifies to the stimulation of redox processes and protein exchange in tissues. Application of the MII-15 preparation is accompanied by an increase in the lactic productivity of cows by 8-10% on the average.     

PURIFICATION AND CHARACTERIZATION OF MUTANASE PRODUCED BY Paenibacillus curdlanolyticus MP-1

Mutanases are enzymes that catalyze hydrolysis of α-1,3-glucosidic bonds in various α-glucans. One of such glucans, mutan, which is synthesized by cariogenic streptococci, is a major virulence factor for induction of dental caries. This means that mutan-degrading enzymes have potential in caries prophylaxis. In this study, we report the purification, characterization, and partial amino acid sequence of extracellular mutanase produced by the MP-1 strain of Paenibacillus curdlanolyticus, bacterium isolated from soil. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme showed a single protein band of molecular mass 134 kD, while native gel filtration chromatography confirmed that the enzyme was a monomer of 142 kD. Mutanase showed a pH optimum in the range from pH 5.5 to 6.5 and a temperature optimum around 40–45°C. It was thermostable up to 45°C, and retained 50% activity after 1 hr at 50°C. The enzyme was fully stable at a pH range of 4 to 10. The enzyme activity was stimulated by the addition of Tween 20, Tween 80, and Ca²⁺, but it was significantly inhibited by Hg²⁺, Ag⁺, and Fe²⁺, and also by p-chloromercuribenzoate, iodoacetamide, and ethylenediamine tetraacetic acid (EDTA). Mutanase preparation preferentially catalyzed the hydrolysis of various streptococcal mutans and fungal α-1,3-glucans. It also showed binding activity to insoluble α-1,3-glucans. The N-terminal amino acid sequence was NH₂-Ala-Gly-Gly-Thr-Asn-Leu-Ala-Leu-Gly-Lys-Asn-Val-Thr-Ala-Ser-Gly-Gln. This sequence indicated an analogy of the enzyme to α-1,3-glucanases from other Paenibacillus and Bacillus species.     

一株新型苯胺蓝降解菌MP-13的代谢特征

[背景]三苯甲烷类染料的广泛使用对我国生态环境构成了极大危害,亟待开发一种经济、高效和环境友好型的染料废水处理技术。目前,利用微生物处理染料废水被认为是一种环境友好型的方法。[目的]通过解析菌株MP-13对苯胺蓝的降解特性,为该菌株在染料废水治理中的应用提供核心理论与技术依据。[方法]从食木白蚁肠道中筛选一株苯胺蓝脱色菌,对其进行16S rRNA基因序列分析鉴定,确定其基本生物学特征,然后通过FTIR、GC/MS等分析手段解析该菌对苯胺蓝的降解特征。[结果]菌株MP-13经鉴定为土白蚁特拉布尔希氏菌(Trabulsiella odontotermitis),该菌对苯胺蓝降解的最适温度、pH和转速分别为35℃、8.0和180 r/min,苯胺蓝浓度为200 mg/L时最大脱色率可达97.3%,且对苯胺蓝的耐受浓度可达1 500 mg/L。此外,FTIR和GC/MS的结果表明苯胺蓝被降解为小分子芳香族化合物。[结论]Trabulsiella odontotemitis MP-13对苯胺蓝有较强降解能力和较高耐受性,可作为染料废水生物修复的潜在菌株资源。     

The TYLCV-tolerant tomato line MP-1 is characterized by superior transformation competence

The tomato line MP-1 excels the cultivars commonly used fortransformation with regard to the speed of regeneration, percentageof transformation and frequency of phenotypically normal transgenicplants. These characteristics, together with its tolerance toTomato Yellow Leaf Curl Virus, make line MP-1 very suitablefor large scale generation of transgenic tomatoes. Key words: Lycopersicon esculentum, transformatio—efficient tomato line     

Oxygen activation and ligand binding by pure heme-octapeptide microperoxidase-8 (MP-8)

The rapid, efficient preparation of pure microperoxidase-8 (MP-8) is described. Ligand binding studies confirm that MP-8 is monomeric in alkaline solution. It is shown that the monomeric MP-8 activates oxygen in a similar manner to that already reported for alkaline hemin, establishing the octapeptide as a possible second generation model for the oxygen activation/insertion reactions of the cytochrome P-450.