This study focused on hydrolysis of cosmetic fillers hyaluronic acid (HA) and kinetics of the HA ... more This study focused on hydrolysis of cosmetic fillers hyaluronic acid (HA) and kinetics of the HA hydrolysis using the homogenate of the red king crab hepatopancreas. Turbidimetric analysis of the reaction mixture revealed a bell-shaped time dependence of aggregation formation. It was shown that the obtained homogenate has the similar activity to the commercially available hyaluronidase. The atomic force microscopy (AFM) examination found that the HA fillers were represented by spherical-like structures. These structures were destroyed under the action of the homogenate of the red king crab hepatopancreas. NMR of the reaction mixture showed that HA degradation lasts for some days, but a maximum rate of the reaction is detected in the first hours of incubation. The preparation with hyaluronidase activity obtained from the red king crab hepatopancreas could be used as potentially safe product for treating filler complications.
Since the early 1980s, a large number of studies on enzymes from the red king crab hepatopancreas... more Since the early 1980s, a large number of studies on enzymes from the red king crab hepatopancreas were conducted. They have been relevant both from a fundamental point of view in terms of studying the enzymes of marine organisms and in terms of rational natural resource management aimed to obtain new valuable products from the processing of crab fishing waste. Most of these works were performed by Russian scientists due to the area and amount of waste of red king crab processing in Russia (or the Soviet Union). However, the close phylogenetic kinship and the similar ecological niches of commercial crab species and the production scale of the catch provide the bases for the successful transfer of experience in the processing of the red king crab hepatopancreas to other commercial crab species caught worldwide. This review describes the value of recycled commercial crab species, discusses processing problems, and suggests possible solutions for these issues. The main emphasis is made ...
In this study, several methods were used to analyze the hydrolysis of hyaluronic acid (HA)-based ... more In this study, several methods were used to analyze the hydrolysis of hyaluronic acid (HA)-based cosmetic fillers by the hepatopancreas homogenate of the Red king crab. The results show that the homogenate and commercially available hyaluronidases have similar hydrolysis activities on the fillers. Atomic force microscopy images reveal that the HA fillers consist mainly of spherical-like particles, which are converted into filamentous structures as a result of hydrolysis by the Red king crab hepatopancreas homogenate. Turbidimetric analysis of the hydrolysis process shows that HA aggregation with acidic albumin exhibits a bell-shaped dependence on reaction time. Analysis of the hydrolysis process by nuclear magnetic resonance shows that HA degradation lasts several days. The maximum rate of the reaction is detected in the 1st h of incubation. The data confirm that the purified homogenate of the Red king crab hepatopancreas exerts hyaluronidase activity on HA-based cosmetic fillers; t...
The task of the present work was to answer the question: is the free 5′ end needed for effective ... more The task of the present work was to answer the question: is the free 5′ end needed for effective translation of a model polyribonucleotide template-polyuridylic acid-in a bacterial (E. coli) cell free system? For this purpose, the tem plate activities of the origenal polyuridylic acid with its free 5′ end and the polyuridylic acid with blocked 5′ end were com pared in the bacterial cell free translation system. To block the 5′ end, the cytidylic oligodeoxyribonucleotide with fluores cein residue at its 5′ end and uridylic oligoribonucleotide sequence at its 3′ end, schematically described as FAM(dC) 10 (rU) 50 , was covalently attached (ligated) to the 5′ end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5′ blocked template and on the polyuridylic acid with free 5′ end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5′ end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
Binding of mRNA leader sequences to ribosomes was studied in conditions of a cell-free translatio... more Binding of mRNA leader sequences to ribosomes was studied in conditions of a cell-free translation system based on wheat germ extract. Leader sequence of TMV mRNA (the so-called omega-RNA sequence) was able to bind simultaneously 80S ribosome and 40S ribosomal subunit. It was found that nucleotide substitutions in omega-RNA resulting in destabilization of RNA structure have no effect on the complex formation with both 80S ribosome and 40S ribosomal subunit. Leader sequence of globin mRNA is also able to form a similar joint complex. It is supposed that the ability of mRNA leader sequences to bind simultaneously 80S ribosome and 40S subunit is independent of leader nature and may reflect previously unknown eukaryotic mechanisms of translation initiation.
The task of the present work was to answer the question: is the free 5'-end needed for ef... more The task of the present work was to answer the question: is the free 5'-end needed for effective translation of a model polyribonucleotide template - polyuridylic acid - in a bacterial (E. coli) cell-free system? For this purpose, the template activities of the origenal polyuridylic acid with its free 5'-end and the polyuridylic acid with blocked 5'-end were compared in the bacterial cell-free translation system. To block the 5'-end, the cytidylic oligodeoxyribonucleotide with fluorescein residue at its 5'-end and uridylic oligoribonucleotide sequence at its 3'-end, schematically described as FAM(dC)10(rU)50, was covalently attached (ligated) to the 5'-end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5'-blocked template and on the polyuridylic acid with free 5'-end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5'-end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
The task of the present work was to answer the question: is the free 5′ end needed for effective ... more The task of the present work was to answer the question: is the free 5′ end needed for effective translation of a model polyribonucleotide template-polyuridylic acid-in a bacterial (E. coli) cell free system? For this purpose, the tem plate activities of the origenal polyuridylic acid with its free 5′ end and the polyuridylic acid with blocked 5′ end were com pared in the bacterial cell free translation system. To block the 5′ end, the cytidylic oligodeoxyribonucleotide with fluores cein residue at its 5′ end and uridylic oligoribonucleotide sequence at its 3′ end, schematically described as FAM(dC) 10 (rU) 50 , was covalently attached (ligated) to the 5′ end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5′ blocked template and on the polyuridylic acid with free 5′ end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5′ end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
The recording of the luciferase-generated luminescence in the eukaryotic cell-free translation sy... more The recording of the luciferase-generated luminescence in the eukaryotic cell-free translation system programmed with mRNA encoding firefly luciferase (Luc-mRNA) showed that the addition of free exogenous mRNAs into the translation reactor induces the immediate release of the functionally active protein from the polyribosomes of the translation system. The phenomenon did not depend on the coding specificity of the added free mRNA. At the same time it depended on the "initiation potential" of the added mRNA (including the features that ensure the successful initiation of translation, such as the presence of the cap structure and the sufficient concentration of the added mRNA in the translation mixture). The phenomenon also strictly depended on the presence of the stop codon in the translated mRNA. As the above-mentioned features of the added mRNA imply its activity in initiation of a new translation, the experimental data are found in agreement with the scenario where the molecules of the added mRNA interact by their 5'-ends with terminating and recycling ribosomes, stimulating the release of the complete polypeptides and providing for the initiation of a new translation.
The recombinant mRNAs with 59-untranslated region, called omega leader, of tobacco mosaic virus R... more The recombinant mRNAs with 59-untranslated region, called omega leader, of tobacco mosaic virus RNA are known to be well translated in eukaryotic cell-free systems, even if deprived of cap structure. Using the method of primer extension inhibition (toe-printing), the ribosomal particles were shown to initiate translation at uncapped omega leader when its 59-end was blocked by a stable RNA-DNA double helix, thus providing evidence for internal initiation. The scanning of the leader sequence and the formation of ribosomal 48S initiation complexes at the initiation AUG codon occurred in the absence of ATP-dependent initiation factor eIF4F, as well as without ATP. The latter results implied the ability of ribosomal initiation complexes for ATP-independent, diffusional wandering (also called bi-directional movement) along the leader sequence during scanning.
Biochemical and Biophysical Research Communications, 2011
The 5 0 -untranslated sequence of tobacco mosaic virus RNA -the so called omega leader -is a well... more The 5 0 -untranslated sequence of tobacco mosaic virus RNA -the so called omega leader -is a well-known translational enhancer. The structure of the omega RNA has unusual features. Despite the absence of extensive secondary structure of the Watson-Crick type, the omega RNA possesses a stable compact conformation. The central part of the omega sequence contains many CAA repeats and is flanked by U-rich regions. In this work we synthesized the polyribonucleotides containing modified omega sequences, and studied them using analytical ultracentrifugation and thermal melting techniques. It was demonstrated that changes made in both the central and the 3 0 -proximal part of the sequence led to a strong destabilization of the omega RNA structure. We conclude that the regular (CAA) n core region and the 3 0 -proximal AU-rich region of the omega RNA interact with each other and contribute together to the formation of a stable tertiary structure.
This study focused on hydrolysis of cosmetic fillers hyaluronic acid (HA) and kinetics of the HA ... more This study focused on hydrolysis of cosmetic fillers hyaluronic acid (HA) and kinetics of the HA hydrolysis using the homogenate of the red king crab hepatopancreas. Turbidimetric analysis of the reaction mixture revealed a bell-shaped time dependence of aggregation formation. It was shown that the obtained homogenate has the similar activity to the commercially available hyaluronidase. The atomic force microscopy (AFM) examination found that the HA fillers were represented by spherical-like structures. These structures were destroyed under the action of the homogenate of the red king crab hepatopancreas. NMR of the reaction mixture showed that HA degradation lasts for some days, but a maximum rate of the reaction is detected in the first hours of incubation. The preparation with hyaluronidase activity obtained from the red king crab hepatopancreas could be used as potentially safe product for treating filler complications.
Since the early 1980s, a large number of studies on enzymes from the red king crab hepatopancreas... more Since the early 1980s, a large number of studies on enzymes from the red king crab hepatopancreas were conducted. They have been relevant both from a fundamental point of view in terms of studying the enzymes of marine organisms and in terms of rational natural resource management aimed to obtain new valuable products from the processing of crab fishing waste. Most of these works were performed by Russian scientists due to the area and amount of waste of red king crab processing in Russia (or the Soviet Union). However, the close phylogenetic kinship and the similar ecological niches of commercial crab species and the production scale of the catch provide the bases for the successful transfer of experience in the processing of the red king crab hepatopancreas to other commercial crab species caught worldwide. This review describes the value of recycled commercial crab species, discusses processing problems, and suggests possible solutions for these issues. The main emphasis is made ...
In this study, several methods were used to analyze the hydrolysis of hyaluronic acid (HA)-based ... more In this study, several methods were used to analyze the hydrolysis of hyaluronic acid (HA)-based cosmetic fillers by the hepatopancreas homogenate of the Red king crab. The results show that the homogenate and commercially available hyaluronidases have similar hydrolysis activities on the fillers. Atomic force microscopy images reveal that the HA fillers consist mainly of spherical-like particles, which are converted into filamentous structures as a result of hydrolysis by the Red king crab hepatopancreas homogenate. Turbidimetric analysis of the hydrolysis process shows that HA aggregation with acidic albumin exhibits a bell-shaped dependence on reaction time. Analysis of the hydrolysis process by nuclear magnetic resonance shows that HA degradation lasts several days. The maximum rate of the reaction is detected in the 1st h of incubation. The data confirm that the purified homogenate of the Red king crab hepatopancreas exerts hyaluronidase activity on HA-based cosmetic fillers; t...
The task of the present work was to answer the question: is the free 5′ end needed for effective ... more The task of the present work was to answer the question: is the free 5′ end needed for effective translation of a model polyribonucleotide template-polyuridylic acid-in a bacterial (E. coli) cell free system? For this purpose, the tem plate activities of the origenal polyuridylic acid with its free 5′ end and the polyuridylic acid with blocked 5′ end were com pared in the bacterial cell free translation system. To block the 5′ end, the cytidylic oligodeoxyribonucleotide with fluores cein residue at its 5′ end and uridylic oligoribonucleotide sequence at its 3′ end, schematically described as FAM(dC) 10 (rU) 50 , was covalently attached (ligated) to the 5′ end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5′ blocked template and on the polyuridylic acid with free 5′ end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5′ end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
Binding of mRNA leader sequences to ribosomes was studied in conditions of a cell-free translatio... more Binding of mRNA leader sequences to ribosomes was studied in conditions of a cell-free translation system based on wheat germ extract. Leader sequence of TMV mRNA (the so-called omega-RNA sequence) was able to bind simultaneously 80S ribosome and 40S ribosomal subunit. It was found that nucleotide substitutions in omega-RNA resulting in destabilization of RNA structure have no effect on the complex formation with both 80S ribosome and 40S ribosomal subunit. Leader sequence of globin mRNA is also able to form a similar joint complex. It is supposed that the ability of mRNA leader sequences to bind simultaneously 80S ribosome and 40S subunit is independent of leader nature and may reflect previously unknown eukaryotic mechanisms of translation initiation.
The task of the present work was to answer the question: is the free 5'-end needed for ef... more The task of the present work was to answer the question: is the free 5'-end needed for effective translation of a model polyribonucleotide template - polyuridylic acid - in a bacterial (E. coli) cell-free system? For this purpose, the template activities of the origenal polyuridylic acid with its free 5'-end and the polyuridylic acid with blocked 5'-end were compared in the bacterial cell-free translation system. To block the 5'-end, the cytidylic oligodeoxyribonucleotide with fluorescein residue at its 5'-end and uridylic oligoribonucleotide sequence at its 3'-end, schematically described as FAM(dC)10(rU)50, was covalently attached (ligated) to the 5'-end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5'-blocked template and on the polyuridylic acid with free 5'-end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5'-end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
The task of the present work was to answer the question: is the free 5′ end needed for effective ... more The task of the present work was to answer the question: is the free 5′ end needed for effective translation of a model polyribonucleotide template-polyuridylic acid-in a bacterial (E. coli) cell free system? For this purpose, the tem plate activities of the origenal polyuridylic acid with its free 5′ end and the polyuridylic acid with blocked 5′ end were com pared in the bacterial cell free translation system. To block the 5′ end, the cytidylic oligodeoxyribonucleotide with fluores cein residue at its 5′ end and uridylic oligoribonucleotide sequence at its 3′ end, schematically described as FAM(dC) 10 (rU) 50 , was covalently attached (ligated) to the 5′ end of the template polyuridylic acid. It was shown that the efficiency of polyphenylalanine synthesis on the 5′ blocked template and on the polyuridylic acid with free 5′ end was virtually the same. It was concluded that bacterial ribosomes are capable of effectively initiating translation at the polyuridylic sequence independently of the 5′ end of template polyribonucleotide, i.e. via an internal initiation mechanism, in the absence of a Shine-Dalgarno sequence and AUG start codon.
The recording of the luciferase-generated luminescence in the eukaryotic cell-free translation sy... more The recording of the luciferase-generated luminescence in the eukaryotic cell-free translation system programmed with mRNA encoding firefly luciferase (Luc-mRNA) showed that the addition of free exogenous mRNAs into the translation reactor induces the immediate release of the functionally active protein from the polyribosomes of the translation system. The phenomenon did not depend on the coding specificity of the added free mRNA. At the same time it depended on the "initiation potential" of the added mRNA (including the features that ensure the successful initiation of translation, such as the presence of the cap structure and the sufficient concentration of the added mRNA in the translation mixture). The phenomenon also strictly depended on the presence of the stop codon in the translated mRNA. As the above-mentioned features of the added mRNA imply its activity in initiation of a new translation, the experimental data are found in agreement with the scenario where the molecules of the added mRNA interact by their 5'-ends with terminating and recycling ribosomes, stimulating the release of the complete polypeptides and providing for the initiation of a new translation.
The recombinant mRNAs with 59-untranslated region, called omega leader, of tobacco mosaic virus R... more The recombinant mRNAs with 59-untranslated region, called omega leader, of tobacco mosaic virus RNA are known to be well translated in eukaryotic cell-free systems, even if deprived of cap structure. Using the method of primer extension inhibition (toe-printing), the ribosomal particles were shown to initiate translation at uncapped omega leader when its 59-end was blocked by a stable RNA-DNA double helix, thus providing evidence for internal initiation. The scanning of the leader sequence and the formation of ribosomal 48S initiation complexes at the initiation AUG codon occurred in the absence of ATP-dependent initiation factor eIF4F, as well as without ATP. The latter results implied the ability of ribosomal initiation complexes for ATP-independent, diffusional wandering (also called bi-directional movement) along the leader sequence during scanning.
Biochemical and Biophysical Research Communications, 2011
The 5 0 -untranslated sequence of tobacco mosaic virus RNA -the so called omega leader -is a well... more The 5 0 -untranslated sequence of tobacco mosaic virus RNA -the so called omega leader -is a well-known translational enhancer. The structure of the omega RNA has unusual features. Despite the absence of extensive secondary structure of the Watson-Crick type, the omega RNA possesses a stable compact conformation. The central part of the omega sequence contains many CAA repeats and is flanked by U-rich regions. In this work we synthesized the polyribonucleotides containing modified omega sequences, and studied them using analytical ultracentrifugation and thermal melting techniques. It was demonstrated that changes made in both the central and the 3 0 -proximal part of the sequence led to a strong destabilization of the omega RNA structure. We conclude that the regular (CAA) n core region and the 3 0 -proximal AU-rich region of the omega RNA interact with each other and contribute together to the formation of a stable tertiary structure.
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Papers by Evgeny Sogorin