Heterogeneous nuclear ribonucleoproteins (hnRNPs) are structurally and functionally distinct proteins containing specific domains and motifs that enable the proteins to bind certain nucleotide sequences, particularly those found in human telomeres. In human malignant cells (HMCs), hnRNP-A1the most studied hnRNPis an abundant multifunctional protein that interacts with telomeric DNA and affects telomerase function. In addition, it is believed that other hnRNPs in HMCs may also be involved in the maintenance of telomere length. Accordingly, these proteins are considered possible participants in the processes associated with HMC immortalization. In our review, we discuss the results of studies on different hnRNPs that may be crucial to solving molecular oncological problems and relevant to further investigations of these proteins in HMCs.

The mitochondrial set of proteins is a dynamic system, crucial for multiple functions of this organelle. Differential expression of genes in various tissues, alternative splicing, post-translational modifications, turnover and spatial dynamics of proteins are the factors that influence mitochondrial proteomes increasing their versatility. A wide range of high-throughput proteomic approaches are extensively used for identification, quantification and functional assessment of human and other mammalian mitochondrial proteins. This article reviews the methods and approaches which can be utilized for achieving one or another specific goal in mitochondrial investigations, and the recent advances in application of proteomics to study the roles of mitochondria in tumorigenesis and cancer progression.

Derivatization of the natural flavonoid dihydroquercetin with p-aminobenzoic acid was carried out in an ethyl acetate/citric buffer biphasic system using laccase from the fungus Trametes hirsuta. The main reaction product yield was ~68 mol %. The product was characterized by 1H NMR, 13C NMR, and liquid chromatography-mass spectroscopy, and its structure was elucidated. The reaction product affected viability of cultured human rhabdomyosarcoma cells (RD cell line) in a dose-dependent manner and, therefore, can be of interest to pharmaceutical industry.

Introduction. Heterogeneous nuclear ribonucleoprotein A1 (hnRNP А1) and other RNA-binding proteins involved in splicing participate in realization of genetic information and can be greatly changed in pathological conditions including tumors. Objective. Proteomic study of hnRNP A1 and other RNA-binding splicing proteins in 10 human malignant and non-malignant cultured cell lines of mesenchymal and epithelial origin. Materials and methods. Two-dimensional gel electrophoresis of adenocarcinomas (LNCaP, DU-145, PC-3, 769-P) and sarcomas (U2-OS, SK-UT-1B, RD) cell lines with following protein identification by matrix-assisted laser desorption ionization mass spectrometry have been carried out. Results. HnRNP А1 has been identified as an abundant protein in all studied malignant cell lines. It has been revealed in lower amount in normal mesenchymal cells compared to malignant cultured cells and achieved undetectable levels in myoblasts after induction of differentiation. Conclusion. High cellular level of hnRNP А1 can suggest high proliferative activity of cells including malignant those. Hence, hnRNP А1 and other RNA-binding splicing proteins hold promise to its further investigation in human transformed cells.

Identification of actin-depolymerizing factor homology (ADF-H) domains in the structures of several related proteins led first to the formation of the ADF/cofilin family, which then expanded to the ADF/cofilin superfamily. This superfamily includes the well-studied cofilin-1 (Cfl-1) and about a dozen different human proteins that interact directly or indirectly with the actin cytoskeleton, provide its remodeling, and alter cell motility. According to some data, Cfl-1 is contained in various human malignant cells (HMCs) and is involved in the formation of malignant properties, including invasiveness, metastatic potential, and resistance to chemotherapeutic drugs. The presence of other ADF/cofilin superfamily proteins in HMCs and their involvement in the regulation of cell motility were discovered with the use of various OMICS technologies. In our review, we discuss the results of the study of Cfl-1 and other ADF/cofilin superfamily proteins, which may be of interest for solving different problems of molecular oncology, as well as for the prospects of further investigations of these proteins in HMCs.

Proteomic technologies have proven to be very effective for detecting biochemical changes in meat products, such as changes in tissue- and species-specific proteins. In the tissues of cattle, pig, horse and camel M. longissimus dorsi both tissue-and species specific proteins were detected using two dimensional electrophoresis. Species-specific isoforms of several muscle proteins were also identified. The identified and described proteins of cattle, pig, horse and camel skeletal muscles (including mass spectra of the tryptic peptides) were added to the national free access database "Muscle organ proteomics". This research has enabled the development of new highly sensitive technologies for meat product quality control against food fraud.

On the basis of the results of proteomic analysis and mass spectrometric identification of human myocardium proteins exhibiting pronounced quantitative changes in the dynamics of prenatal cardiogenesis, changes in the expression level of proteins of three families (mitochondrial, contractile, and heat shock) have been identified. The complex of human myocardium mitochondrial proteins (for example, α and β isoforms of ATP synthase, aconitase 2, creatine phosphokinase M-subunit, and 60-kDa heat shock protein) largely finishes its development according to the adult type by developmental week 24. The formation of the protein composition of human myocardium contractile structures (for example, desmin, myosin regulatory light chain 2, fetal ventricular essential isoform 1, canonical α-tropomyosin, and fetal isoform 6) reflects the initial stage of myofibril development until developmental week 8 (replacement of fetal isoforms of contractile proteins with adult ones with the involvement of the phosphorylated isoform of 27-kDa heat shock protein), the stage of their qualitative and quantitative structuring by developmental weeks 20–24, and the final formation of the adult phenotype of contractile structures by 2 years of life.

Modern biochemistry or biochemistry of 21th century is developing in many traditional fields: investigations of proteins, nucleic acids, lipids, carbohydrates, metabolites and metabolic processes, etc, however it has gained new features that based on success of Human Genome Project and on application of high performance technologies (post-genomic technologies). As a result, at the turn of the 21st century some new scientific disciplines, named “OMICS” (proteomics, transcriptomics, lipidomics, glycomics, metabolomics, etc) were created through advances in the biochemistry. Currently “OMICS” and post-genomic technologies are involved in mainstream of cancer research, including studies of human malignant cells. Since malignant tumors consist of heterogeneous cancer cells, the cultured cell lines have some advantages over biopsy simples for studies aimed at understanding the molecular basis of carcinogenesis. The malignant human cell lines differ considerably in their origin from tissues or organs, and therefore they vary widely in the differentiation characteristics and the gene expression patterns. Thus, the use of malignant cell lines ex vivo (as models of human cancers) is an important trend in search for key molecules of carcinogenesis in experimental oncology

Splicing factor, proline- and glutamine-rich protein (SFPQ), was identified in eight human cultivated cell lines by proteomic approaches. The cell proteins have been separated by means of two-dimensional gel electrophoresis in two modifications and identified by matrix-assisted laser desorption ionization mass spectrometry with further tandem mass spectrometry. The analysis of proteins from three human sarcomas cell lines (RD, U-2 OS and SK-UT-1B), three human renal adenocarcinomas cell lines (A-498, 769-P and OKP-GS), and two prostate adenocarcinomas cell lines (DU-145 and PC-3) revealed several electrophoretic isoforms of SFPQ protein. Differences between theoretical and experimental molecular masses and isoelectric points of SFPQ protein have been observed. Detailed investigation of SFPQ peptides by tandem mass spectrometry has detected new phosphorylation state of threonine residue in 168 position of SFPQ isoform in rhabdomyosarcoma cell line. Furthermore, SFPQ has not been identified during proteomic study of several nonmalignant cell lines, including cultured human mesenchymal stromal cells and myoblasts. However, SFPQ has been found in all malignant cell lines in high quantity. In particular, its fractions are abundant in sarcomas cell lines as opposed to nonmalignant mesenchymal cells. It is assumed that high quantity of SFPQ in sarcomas cell lines may affect tumorigenesis.

The article presents the results of autolytic processes impact on the protein-peptide profile of Bos taurus and Sus scrofa cardiac muscle and aorta. The results of tissue-specific protein identification are also presented as well as the effect of autolysis. Apolipoprotein A-1 involved in the formation of high-density lipoproteins, peroxiredoxin-1 involved in the suppression of oxidative stress, galectin-1 induced apoptosis of T-lymphocytes, as well as number of heat shock proteins with molecular weight less than 30 kDa were identified in Sus scrofa aorta tissue. It was discovered that functional proteins with molecular weight less than 30 kDa are retained during the freezing process, but destroyed under the action of autolytic enzymes. This work was supported by the Russian Science Foundation (project No. 16–16–10073).

Proteomic technologies in the modern laboratory practice proved to be very efficient to reveal biochemical changes in meat products, such as changes in heat-resistant and species-specific proteins that have the ability to become the relevant bio-markers. Several tissue-specific proteins were identified in the work under review using proteomic technologies in tested samples of meat and in specially manufactured sausage products that may be used as individual biomarkers to verify conformity of meat products to the alleged composition. Also, individual non-muscle proteins (soya and chicken protein) were determined in test samples of meat products apart from species-specific muscle proteins that may act as functional ingredients used in cooking process. Overall, total of more than 200 protein fractions were identified in the completed studies by the mass spectrometry method which are described in this review in part. The results obtained will be used to draft the procedure for quantitative evaluation of the meat component content in structureless cooked products (cooked sausages) as well as to draw proteomic protein charts of the native meat stock used to manufacture goods as per GOST (State Standard). Studies conducted in the range of this discipline will help to formulate and considerably develop approaches to identify and evaluate protein markers of quality, functionality and safety of meat for processing and processed meat products.

The skeletal muscle protein troponin I (TnI) has been characterized as a potential thermally stable and species-specific biomarker of mammalian muscle tissues in raw meat and meat products. This study proposed a technique for the quantification of TnI comprising protein extraction and sandwich enzyme-linked immunosorbent assay (ELISA). The technique is characterized by a TnI detection limit of 4.8 ng/ml with quantifiable concentrations ranging from 8.7 to 52 ng/ml. The method was shown to be suitable for detection of TnI in mammalian (beef, pork, lamb, and horse) meat but not in poultry (chicken, turkey, and duck) meat. In particular, the TnI content in beef was 0.40 3 ± 0.058 mg/g of wet tissue. The TnI estimations obtained for the pork and beef samples using ELISA were comparable to the proteomic analysis results. Thus, the quantitative study of TnI can be a convenient way to assess the mammalian muscle tissue content of various meat products.

The review briefly summarizes the data on the development of proteomic technologies that became actively used in studies of the muscular proteins of farm animals used in the meat industry in 2006-2013. It has been noted that the main research trends are connected with the detection of changes in muscle proteins during post-mortem autolysis and the search for species-specific and other protein biomarkers. Particular publications regarding the development of methods based on proteomic technologies for monitoring the state of muscle proteins are considered. According to the analyzed data, we can conclude that the field is promising for the solution of a number of pressing problems in.applied biochemistry.

A new biotest system was developed based on highly proliferating human cell cultures (lines LNCaP and PC-3). With the help of this system, two known synthetic polyamines—α-difluoromethylornithine (DFMO) and methylglioxalbis(guanylhydrason) (MGBG)—as well as four new synthetic analogues difenylcontaining amines (DFCA-1-DFCA-4) with molecular weights of 725.5 (DFCA-1), 755.5 (DFCA-2), 655.5 (DFCA-3), and 681.5 Da (DFCA-4) were tested. In this biotest system, DFMO (0.1–400 μM) did not reveal functional activity, whereas for MGBG a cytotoxic effect was registered (100–200 μM). DFCA-1, DFCA-2, and DFCA-4 had a similar effect at concentrations of 10 μM and higher; DFCA-3, at a concentration of 50 μM and higher. Thus, DFCA-1 has a higher level of antiproliferating activity and may be considered as the most potent cytostatic agent.

Transgelins, which have been studied by proteomic methods for more than ten years as tumor-associated proteins, are the members of calponin family, since calponin homology domains are present in their amino acid sequences and there are other typical structural features. According to the available information there is a specific tissue distribution of these proteins. Three closely related genes which coded transgelins are revealed in the human genome. Single nucleotide substitutions and the expression peculiarities of these genes can be the reasons for existence of several isoforms of three main transgelins. Different manifestations of biochemical polymorphism of transgelins are detected by now. Transgelin (or SM22-alpha) is well known as a biomarker of smooth muscle cell differentiation. It was revealed that transgelin acts as repressor of MMP9 gene, coding matrix metalloproteinase-9. At the same time the oppositely directed changes in the content of transgelin proteins were found in tissue samples of malignant tumors of the respiratory, digestive and urogenital systems, as well as in the cultivated tumor cell using proteomic technologies. Correspondingly, according to some authors transgelins can be potential biomarkers of malignant neoplasms, but in the opinion the others - transgelins are suppressors of tumor growth. In this review, we consider possible reasons for such differences and discuss the prospects of solution of the existing contradictions.

Proteomic analysis of proteins in samples of raw meat, two kinds of cooked sausages (Doctorskaya and Lyubitelskaya), and also specially made functional meat products was performed. The mass spectrometry methods allowed to identify more than 200 protein fractions, including 51 in beef samples and 108 in pork samples. It is noted that the proteomic profiles of the studied samples have a certain similarity caused by the presence a number of tissue-specific sarcomeric proteins (troponins, muscle isoform of myosin light chain, -tropomyosin, desmin). In the samples of meat products some species-specific muscle proteins as well as individual proteins of nonmuscular origin are revealed which can be considered as functional ingredients. It is shown that in processes of posthumous autolysis and production of meat products some protein are exposed to specific and nonspecific proteolytic degradation. The received results in the summarized form are included in the information database Muscle organs proteomics, version 2013 (http://mp.inbi.ras.ru).

На основе быстро пролиферирующих культивируемых клеток человека (линии LNCaP и РС-3) разработана новая биотест-система, с помощью которой охарактеризованы два известных синтетических полиамина – -дифторметилорнитин (ДФМО) и метилглиоксальбис(гуанилгидразон) (МГБГ), а также 4 новых синтетических аналога – дифенил-содержащие амины (ДФСА-1–ДФСА-4) с молекулярными массами (Да): ДФСА-1 – 725.5; ДФСА-2 – 755.5; ДФСА-3 – 655.5; ДФСА-4 – 681.5. В этой биотест-системе ДФМО (0.1–400 мкМ) не проявил функциональной активности, тогда как у МГБГ был зарегистрирован цитостатический эффект (100–200 мкМ). ДФСА-1, -2 и -4 оказывали подобный эффект при концентрациях 10 мкМ и выше, а ДФСА-3 – при концентрации 50 мкМ и выше. При этом ДФСА-1 с наибольшим антипролиферативным действием представляется особенно перспективным цитостатическим агентом.

This review considers the major features of human proteins AGR2 and ERp57/GRP58 and of other members of the protein disulfide isomerase (PDI) family. The ability of both AGR2 and ERp57/GRP58 to catalyze the formation of disulfide bonds in proteins is the parameter most important for assigning them to a PDI family. Moreover, these proteins and also other members of the PDI family have specific structural features (thioredoxin-like domains, special C-terminal motifs characteristic for proteins localized in the endoplasmic reticulum, etc.) that are necessary for their assignment to a PDI family. Data demonstrating the role of these two proteins in carcinogenesis are analyzed. Special attention is given to data indicating the presence of biomarker features in AGR2 and ERp57/GRP58. It is now thought that there is sufficient reason for studies of AGR2 and ERp57/GRP58 for possible use of these proteins in diagnosis of tumors. There are also prospects for studies on AGR2 and ERp57/GRP58 leading to developments in chemotherapy. Thus, we suppose that further studies on different members of the PDI family using modern postgenomic technologies will broaden current concepts about functions of these proteins, and this will be helpful for solution of urgent biomedical problems.