The Analysis of OmpA and Rz/Rz1 of Lytic Bacteriophage from Surabaya, Indonesia

A good strategy to conquer the Escherichia coli-cause food-borne disease could be bacteriophages. Porins are a type of β-barrel proteins with diffuse channels and OmpA, which has a role in hydrophilic transport, is the most frequent porin in E. coli; it was also chosen as the potential receptor of the phage. And the Rz/Rz1 was engaged in the breakup of the host bacterial external membrane. This study aimed to analyze the amino acid of OmpA and Rz/Rz1 of lytic bacteriophage from Surabaya, Indonesia. This study employed a sample of 8 bacteriophages from the previous study. The OmpA analysis method was mass spectrometry. Rz/Rz1 was analyzed using PCR, DNA sequencing, Expasy Translation, and Expasy ProtParam. The result obtained 10% to 29% sequence coverage of OmpA, carrying the ligand-binding site. The Rz/Rz1 gene shares a high percentage of 97.04% to 98.89% identities with the Siphoviridae isolate ctTwQ4, partial genome, and Myoviridae isolate cthRA4, partial genome. The Mann–Whitney statistical tests indicate the significant differences between Alanine, Aspartate, Glycine, Proline, Serine ( p = 0.011 ), Asparagine, Cysteine ( p = 0.009 ), Isoleucine ( p = 0.043 ), Lysine ( p = 0.034 ), Methionine ( p = 0.001 ), Threonine ( p = 0.018 ), and Tryptophan ( p = 0.007 ) of OmpA and Rz/Rz1. The conclusion obtained from this study is the fact that OmpA acts as Phage 1, Phage 2, Phage 3, Phage 5, and Phage 6 receptors for its peptide composition comprising the ligand binding site, and Rz/Rz1 participates in host bacteria lysis.

Protein-peptide composition in the lungs of rats with hyperhomocysteinemia

The accumulated data indicate that a high level of homocysteine may be a central pathogenetic factor of chronic obstructive pulmonary disease. In this study, we investigated the effect of hyperhomocysteinemia on protein homeostasis in the rat lungs. The level of proteins, peptides, total proteolytic activity, as well as protein-peptide composition, were evaluated. Hyperhomocysteinemia was induced by daily intragastric administration of DL-homocysteine thiolactone (100 mg·kg-1 of body weight) to albino non-linear male rats for 28 days. Twelve hours after the last administration, the rats were sacrificed and the lungs were harvested. Our findings showed that hyperhomocysteinemia caused the disturbances in the protein homeostasis in the lungs that are manifested by a decrease in the level of proteins in the young and old animals and an increase in the level of peptides in the rats of all studied groups. We found a change in the protein composition in the lung of HM rats - a decrease in the level of proteins with a molecular weight of 50 kDa to 100 kDa simultaneously with an increase in the level of proteins with a molecular weight of less than 50 kDa. Despite the fact that the peptide profile was the same in both control animals and HM animals, the level of individual peptide fractions increased significantly in the rats with HM. Obtained data could contribute to explain, at least in part, the mechanisms involved in the pathogenesis of lung damage in hyperhomocysteinemia.

Synthesis and Analysis of Properties of an Immunogenic Fragment from NS4A Polypeptide of Hepatitis C Virus

Abstract— The work is aimed at the synthesis and analysis from NS4A of hepatitis C virus (HCV) antigen peptide fragment that contains a conserved B-cell and T-helper epitopes. The 24-mer peptide VIVGRIILSGRPAVIPDREVLYRK-NH2, which contains the main immunogenic site 24–46 of HCV NS4A antigen (corresponding to the 1681–1703 amino acid residues of the HCV polypeptide), subtype 1b, has been prepared via solid-phase synthesis according to the Fmoc-protocol. Particles with diameters of 73 ± 10 nm (30%) and 236 ± 5 nm (70%) have been detected in the water solution of the highly purified peptide (0.5 mg/mL) by dynamic light scattering. The polydispersity index of 0.377 ± 0.012 implies the existence of heterogeneity because of the aggregation of the peptide molecules. The ζ-potential of the peptide aggregates has been determined as 7.0 ± 0.5 mV by means of electrophoretic light scattering. These data confirm the possibility for the development of a nanoscale liposome form of the peptide preparation. Immunoreactivity of the synthesized highly purified peptide has been studied with the use of blood sera of patients with chronic hepatitis C. Antipeptide immunoglobulins G have been detected in 41.7% of serum samples. Thus, this peptide has been shown to reproduce at least one B-epitope, to which antibodies are raised during natural HCV infection. The synthesized 24-mer peptide is a promising candidate for further research and for use as a potential immunogen for the design of a nanoscale therapeutic immunogenic liposomal peptide composition with synthetic lipids as an adjuvant.

Amphiphilic RGD and GHK peptides synergistically enhance liposomal delivery into cancer and endothelial cells

This study reveals enhanced cancer-targeting properties of peptide composition consisting of RGD and GHK recognized as important cell adhesion factor and pleiotropic modulator of cellular functions, respectively. C12-GGRGD-NH2 and C12-GGGHK-NH2...

Exploring Potential Bioactive Peptides in Fermented Bactrian Camel’s Milk and Mare’s Milk Made by Mongolian Nomads

To date, bioactive proteins and peptides from minor livestock milks and their fermented products have been scarcely reported. In Mongolia, nomads are commonly rearing five livestock animal species (i.e., cow, camel, goat, horse, and sheep) for milking and other purposes. In this study, we analyzed the peptide composition in fermented milks of Bactrian camels (Camelus bactrianus) and horses, produced by Mongolian nomads for self-consumption. Peptides from skimmed fermented milks were separated by ultrafiltration and reverse-phase high-performance liquid chromatography. Then, their amino acid sequences were determined by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. Consequently, eleven peptides were identified in the fermented camel’s milk including four from β-casein (β-CN), three from αs1-CN, and two from both κ-CN and lactophorin. On the other hand, twenty-four peptides were identified in the fermented mare’s milk including nineteen from β-CN, three from αs1-CN, and one from both κ-CN and αs2-CN. According to previous reports on the bioactivities of milk-derived peptides, antibacterial and antihypertensive activities were promising in both the fermented camel’s milk and mare’s milk. In addition, potential antioxidant activity was conjectured in the fermented camel’s milk. Further investigations are currently needed to clarify the potential role of immunomodulatory peptides in the two fermented milks.

A combined microphysiological-computational omics approach in dietary protein evaluation

Food security is under increased pressure due to the ever-growing world population. To tackle this, alternative protein sources need to be evaluated for nutritional value, which requires information on digesta peptide composition in comparison to established protein sources and coupling to biological parameters. Here, a combined experimental and computational approach is presented, which compared seventeen protein sources with cow’s whey protein concentrate (WPC) as the benchmark. In vitro digestion of proteins was followed by proteomics analysis and statistical model-based clustering. Information on digesta peptide composition resulted in 3 cluster groups, primarily driven by the peptide overlap with the benchmark protein WPC. Functional protein data was then incorporated in the computational model after evaluating the effects of eighteen protein digests on intestinal barrier integrity, viability, brush border enzyme activity, and immune parameters using a bioengineered intestine as microphysiological gut system. This resulted in 6 cluster groups. Biological clustering was driven by viability, brush border enzyme activity, and significant differences in immune parameters. Finally, a combination of proteomic and biological efficacy data resulted in 5 clusters groups, driven by a combination of digesta peptide composition and biological effects. The key finding of our holistic approach is that protein source (animal, plant or alternative derived) is not a driving force behind the delivery of bioactive peptides and their biological efficacy.

Worldwide experience of the therapeutic use of the human placental hydrolytes

Here we present the results of a systematic analysis of publications on the clinical and the experimental pharmacology of human placental hydrolyzates (HPH). Searches were performed in the PUBMED, ELIBRARY and RSL databases. The results of the analyses of the peptide composition of HPH allowed us to formulate a number of previously unknown molecular mechanisms of their action. In the article we examine the effects of HPH in the therapy of liver diseases, atopic dermatitis, herpetic infection, viral hepatitis, joint diseases, iron overload, chronic fatigue syndrome and consider the general regenerative abilities of the HPH.

PSV-29 Changes in the PSE and DFD porcine muscle proteome during autolysis

Despite reducing of pale, soft, exudative (PSE) pork share, its amount is 45% in Russia and 65% in Belarus of the total. Dark, firm, dry (DFD) pork occurrence increases to 20% in Russia and 30% in Belarus. Protein changes in normal (NOR), PSE and (DFD) pork muscle during 5 days ageing were studied. Were found differences in their protein profiles and potential protein markers of such raw materials were identified. Mainly in range 600–1000 Da and up to 450 Da. Analysis of the peptide composition showed two myosin light chain isoforms (MLC) (Mw/pI): MLC-like protein — 21.8/5.00 and phosphorylated MLC f -21.0/4.90, in the sample. One of which is fast, the other is slow MLC isoform. Increase in the number of fragments was noted of such proteins as pyruvatekinase, α-enolase, muscle creatine phosphokinase and GPDH, adenylatekinase, αB-crystallin. A correlation between muscle proteins breakdown (the troponin group/troponin family) and the characteristics of meat was found. In NOR samples, troponin I decomposed quite clearly. The most obvious autolysis marker is the appearance and increase of three additional fragments of fast skeletal muscle troponin T, which differ in molecular weight and pI. According to the tryptic peptides mass spectra, the C- and N-molecule terminal parts peptides are no longer detected sequentially. Moreover, in DFD samples, they are most significantly split into shorter peptides. Appearance of short N-terminal GPDH 21 kDa fragment on the 5th day of autolysis is PSE specific marker. Totally 12, 14 and 18 specific peptides were identified in chilled and 18, 32 and 38 on 5th day of autolysis in PSE, NOR and DFD pork respectively. DFD meat can be considered a source of natural specific low-molecular peptides, specifically to produce meat products with functional properties. (Research was supported by the Russian Science Foundation, project 16-16-10073)