Alexander Zhyvoloup | University College London (original) (raw)
Papers by Alexander Zhyvoloup
PLOS ONE
The development of inhibitors of islet amyloid formation is important as pancreatic amyloid depos... more The development of inhibitors of islet amyloid formation is important as pancreatic amyloid deposition contributes to type-2 diabetes and islet transplant failure. The Alzheimer's Aβ peptide and human amylin (h-amylin), the polypeptide responsible for amyloid formation in type-2 diabetes, share common physio-chemical features and some inhibitors of Aβ also inhibit amyloid formation by h-amylin and vice versa. Thus, a popular and potentially useful strategy to find lead compounds for anti-amylin amyloid agents is to examine compounds that have effects on Aβ amyloid formation. The triphenylmethane dye, brilliant blue G (BBG, Sodium;3-[[4-[(E)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-N-ethyl-3-methylanilino]methyl] benzenesulfonate) has been shown to modulate Aβ amyloid formation and inhibit Aβ induced toxicity. However, the effects of BBG on h-amylin have not been examined, although other triphenylmethane derivatives inhibit h-amylin amyloid formation. The compound has only a modest impact on h-amylin amyloid formation unless it is added in significant excess. BBG also remodels preformed h-amylin amyloid fibrils if added in excess, however BBG has no significant effect on h-amylin induced toxicity towards cultured β-cells or cultured CHO-T cells except at high concentrations. BBG is shown to interfere with standard thioflavin-T assays of h-amylin amyloid formation and disaggregation, highlighting the difficulty of interpreting such experiments in the absence of other measurements. BBG also interferes with ANS based assays of h-amylin amyloid formation. The work highlights the differences between inhibition of Aβ and h-amylin amyloid formation, illustrates the limitation of using Aβ inhibitors as leads for h-amylin amyloid inhibitors, and reinforces the difficulties in interpreting dye binding assays of amyloid formation.
Biochemical and Biophysical Research Communications
Aurora A is a cell cycle protein kinase implicated in multiple human cancers, and several Aurora ... more Aurora A is a cell cycle protein kinase implicated in multiple human cancers, and several Aurora A-specific kinase inhibitors have progressed into clinical trials. In this study, we report structural and cellular analysis of a novel biochemical mode of Aurora A inhibition, which occurs through reversible covalent interaction with the universal metabolic integrator coenzyme A (CoA). Mechanistically, the CoA 3'-phospho ADP moiety interacts with Thr 217, an Aurora A selectivity filter, which permits the formation of an unprecedented covalent bond with Cys 290 in the kinase activation segment, lying some 15 A away. CoA modification (CoAlation) of endogenous Aurora A is rapidly induced by oxidative stresses at Cys 290 in human cells, and microinjection of CoA into mouse embryos perturbs meitoic spindle formation and chromosome alignment. Aurora A regulation by CoA reveals how targeting of Aurora A might be accomplished in the future by development of a 'double-anchored' coval...
The Biochemical journal, Jan 6, 2018
In all living organisms, coenzyme A (CoA) is an essential cofactor with a unique design allowing ... more In all living organisms, coenzyme A (CoA) is an essential cofactor with a unique design allowing it to function as an acyl group carrier and a carbonyl-activating group in diverse biochemical reactions. It is synthesized in a highly conserved process in prokaryotes and eukaryotes that requires pantothenic acid (vitamin B5), cysteine and ATP. CoA and its thioester derivatives are involved in major metabolic pathways, allosteric interactions and the regulation of gene expression. A novel unconventional function of CoA in redox regulation has been recently discovered in mammalian cells and termed protein CoAlation. Here, we report for the first time that protein CoAlation occurs at a background level in exponentially growing bacteria and is strongly induced in response to oxidizing agents and metabolic stress. Over 12% of gene products were shown to be CoAlated in response to diamide-induced stress CoAlation of glyceraldehyde-3-phosphate dehydrogenase was found to inhibit its enzymatic...
PLOS Pathogens
HIV-1 integrates more frequently into transcribed genes, however the biological significance of H... more HIV-1 integrates more frequently into transcribed genes, however the biological significance of HIV-1 integration targeting has remained elusive. Using a selective high-throughput chemical screen, we discovered that the cardiac glycoside digoxin inhibits wild-type HIV-1 infection more potently than HIV-1 bearing a single point mutation (N74D) in the capsid protein. We confirmed that digoxin repressed viral gene expression by targeting the cellular Na + /K + ATPase, but this did not explain its selectivity. Parallel RNAseq and integration mapping in infected cells demonstrated that digoxin inhibited expression of genes involved in Tcell activation and cell metabolism. Analysis of >400,000 unique integration sites showed that WT virus integrated more frequently than N74D mutant within or near genes susceptible to repression by digoxin and involved in T-cell activation and cell metabolism. Two main gene networks down-regulated by the drug were CD40L and CD38. Blocking CD40L by neutralizing antibodies selectively inhibited WT virus infection, phenocopying digoxin. Thus the selectivity of digoxin depends on a combination of integration targeting and repression of specific gene networks. The drug unmasked a functional connection between HIV-1 integration and T-cell activation. Our results suggest that HIV-1 evolved integration site selection to couple its early gene expression with the status of target CD4+ T-cells, which may affect latency and viral reactivation.
The Biochemical journal, Jul 24, 2017
Coenzyme A (CoA) is an obligatory cofactor in all branches of life. CoA and its derivatives are i... more Coenzyme A (CoA) is an obligatory cofactor in all branches of life. CoA and its derivatives are involved in major metabolic pathways, allosteric interactions and the regulation of gene expression. Abnormal biosynthesis and homeostasis of CoA and its derivatives have been associated with various human pathologies, including cancer, diabetes and neurodegeneration. Using an anti-CoA monoclonal antibody and mass spectrometry we identified a wide range of cellular proteins which are modified by covalent attachment of CoA to cysteine thiols (CoAlation). We show that protein CoAlation is a reversible post-translational modification that is induced in mammalian cells and tissues by oxidising agents and metabolic stress. A number of key cellular enzymes were found to be CoAlated in vitro and in vivo in ways that modified their activities. Our study reveals that protein CoAlation is a widespread post-translational modification which may play an important role in redox regulation under physiol...
Retrovirology, 2016
Question: Foamy viruses (FV), and in particular PFV, have emerged in recent years as attractive g... more Question: Foamy viruses (FV), and in particular PFV, have emerged in recent years as attractive gene therapy vector candidates. Since the lack of knowledge on molecular events in FV replication is a major hurdle for broader usage of foamy virus vectors, we aimed at elucidating PFV biology by investigating interactions of its capsid protein, Gag, with host cell components. Methodology and result: To this end, we identified members of the mammalian PLK family as PFV Gag interactants in a commercial yeasttwo-hybrid (Y2H) screen and validated these results in detailed Y2H experiments for PLK1-3. In the yeast system, the intact PLK kinase and substrate recognition motifs were required for interactions with PFV Gag, in which a unique S 224-T-P 226 motif served as a PLK binding determinant. PFV Gag mutants harbouring alanine substitutions of STP residues (iSTP) or phosphomimetic mutations of the T 225 (pmSTP) failed to interact with PLK1-3 in yeast. These findings were corroborated by colocalization studies of ectopically expressed, fluorescently tagged proteins in mammalian cells, where mCherry-tagged PFV Gag was able to recruit eGFP-tagged PLK1 and 2 to condensed mitotic chromatin in an STP motif-dependent manner. When characterizing PFV virions containing wild type or STP mutant Gag proteins, we observed that the mutations did not interfere with particle assembly, release or reverse transcription, but led to a 70 % titer reduction relative to wild type in single-round infection experiments. These replication defects became more prominent in the replication-competent PFV context. Therefore, the lack of Gag STP mutant interaction with PLK proteins upon viral entry into host cells was likely underlying this replication deficit. This hypothesis was strengthened by the finding that enzymatic PLK inhibition in host cells during transduction with wild type PFV mimicked the replication phenotype of PFV STP mutants. In addition to the overall reduced infectivity of the mutants, we also observed that the STP mutations in particle-associated Gag lead to differential sensitivity to integrase inhibition by dolutegravir and resulted in decreased integration efficiency. Conclusions: Taken together, our results demonstrate that PLK proteins influence PFV replication by virtue of their interaction with the Gag protein, ensuring timely and efficient transduction. O2 A novel entry/uncoating assay reveals the presence of at least two species of viral capsids during synchronized HIV-1 infection
Euphytica, 2000
The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan a... more The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan and Taiwan was examined with RAPD-PCR (Random Amplified Polymorphic DNA Polymerase Chain Reaction) markers. Out of the 50 primers screened, 17 primers generated 58 polymorphic and reproducible bands. A minimum of 3 primers was sufficient to distinguish all the 27 accessions studied. The Shannon's index used to partition diversity into inter-and intra-group, revealed that 71 percent of variability resided within groups and 29 percent between groups. Diversity was greatest within the Korean group followed by Taiwan and Japan. The relatively high diversity observed in Korea might reflect the larger genetic base of its plantations while the low diversity in Japan could be explained by the long and intensive tea selection programme in this country. A dendrogram based on the UPGMA-link method using Jaccard's distances and multivariate Factorial correspondence analysis clustered the tea accessions into two main groups, regrouping the Taiwan cultivars on the one side and the Korean and Japanese accessions on the other side. This suggests that the Taiwan tea studied here may have a different origin from that of Korea and Japan.
Molekuliarnaia genetika, mikrobiologiia i virusologiia
Biopolymers and Cell
Розроблено систему оцінки та порівняно in vivo регуляторну здатність рибосомних білків L10. Кліти... more Розроблено систему оцінки та порівняно in vivo регуляторну здатність рибосомних білків L10. Клітини Escherichia coli котрансформуються плазмідами-продуцентами LlO та плазмідою з репортерним геном rplJ'-IacZ. Регуляторна здатність оцінюється за ефективністю пригнічування білком LlO експресії репортерного гену.
Molekuliarnaia genetika, mikrobiologiia i virusologiia
The recombinant plasmids pUC and pNM481 were constructed. The efficiency of P1ac promoter of the ... more The recombinant plasmids pUC and pNM481 were constructed. The efficiency of P1ac promoter of the vector plasmid pUC and PL10. supporting the transcription of rplJL-rpoBC-operons of Escherichia coli was compared. It was found that the stronger promoter P1ac, due to convergent transcription directed by the promoter, makes possible the cloning on the multicopy plasmid pUC of the DNA fragment (the gene rplJ controlled by its own promoter PL10), coding for the synthesis of the regulatory ribosomal protein L10.
Biopolymers and Cell
Короткі повідомлення УДК 677.21 Α. Μ. Живолуп, Є. Б. Патон МОЖЛИВІСТЬ ГЕТЕРОЛОГІЧНОЇ РЕГУЛЯЦІЇ ЕК... more Короткі повідомлення УДК 677.21 Α. Μ. Живолуп, Є. Б. Патон МОЖЛИВІСТЬ ГЕТЕРОЛОГІЧНОЇ РЕГУЛЯЦІЇ ЕКСПРЕСІЇ ГЕНІВ ОПЕРОНУ гр1JL KLEBSIELLA PNEUMONIAE БІЛКОМ L10 ESCHERICHIA COLI Показано можливість досягнення високої ефективності трансформації К. pneumoniae методом електропорації. Виявлено регуляторну здатність білків LlO Е. соїі з нативною та зміненою первинною структурою стосовно генів оперону rplJL К. pneumoniae.
Experimental oncology, 2006
To clone, express in baculovirus expression system and purify SSX2 tumor antigen and to use it fo... more To clone, express in baculovirus expression system and purify SSX2 tumor antigen and to use it for screening of blood serum of melanoma patients. Cloning and expression of SSX2 antigen in Bac-to-Bac baculovirus expression system as His-tag fusion protein, expression of recombinant SSX2 in insect cells with following purification by affinity chromatography on Ni-NTA agarose, ELISA of blood serum of melanoma patients (n = 29) and healthy donors (n = 27) were used. SSX2 was cloned in baculovirus expression system, expressed, purified using affinity chromatography and used in ELISA as antigen. Comparative analysis of blood serum of melanoma patients and healthy donors revealed higher level of SSX2-positivity of blood serum from the patients with cancer. SSX2 antigen expressed in baculovirus expression system can be used for serological analysis of blood serum of oncological patients.
Genetika, 1997
Regulatory interaction of the L10 protein and translation operator of L10 mRNA was studied in viv... more Regulatory interaction of the L10 protein and translation operator of L10 mRNA was studied in vivo using a double-plasmid system. Feedback regulation of rplJ gene expression in Thermotoga maritima was proved, and the possibility of feedback cross-regulation was demonstrated in a heterologous system containing T. maritima and enterobacterial components.
PLOS ONE
The development of inhibitors of islet amyloid formation is important as pancreatic amyloid depos... more The development of inhibitors of islet amyloid formation is important as pancreatic amyloid deposition contributes to type-2 diabetes and islet transplant failure. The Alzheimer's Aβ peptide and human amylin (h-amylin), the polypeptide responsible for amyloid formation in type-2 diabetes, share common physio-chemical features and some inhibitors of Aβ also inhibit amyloid formation by h-amylin and vice versa. Thus, a popular and potentially useful strategy to find lead compounds for anti-amylin amyloid agents is to examine compounds that have effects on Aβ amyloid formation. The triphenylmethane dye, brilliant blue G (BBG, Sodium;3-[[4-[(E)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-N-ethyl-3-methylanilino]methyl] benzenesulfonate) has been shown to modulate Aβ amyloid formation and inhibit Aβ induced toxicity. However, the effects of BBG on h-amylin have not been examined, although other triphenylmethane derivatives inhibit h-amylin amyloid formation. The compound has only a modest impact on h-amylin amyloid formation unless it is added in significant excess. BBG also remodels preformed h-amylin amyloid fibrils if added in excess, however BBG has no significant effect on h-amylin induced toxicity towards cultured β-cells or cultured CHO-T cells except at high concentrations. BBG is shown to interfere with standard thioflavin-T assays of h-amylin amyloid formation and disaggregation, highlighting the difficulty of interpreting such experiments in the absence of other measurements. BBG also interferes with ANS based assays of h-amylin amyloid formation. The work highlights the differences between inhibition of Aβ and h-amylin amyloid formation, illustrates the limitation of using Aβ inhibitors as leads for h-amylin amyloid inhibitors, and reinforces the difficulties in interpreting dye binding assays of amyloid formation.
Biochemical and Biophysical Research Communications
Aurora A is a cell cycle protein kinase implicated in multiple human cancers, and several Aurora ... more Aurora A is a cell cycle protein kinase implicated in multiple human cancers, and several Aurora A-specific kinase inhibitors have progressed into clinical trials. In this study, we report structural and cellular analysis of a novel biochemical mode of Aurora A inhibition, which occurs through reversible covalent interaction with the universal metabolic integrator coenzyme A (CoA). Mechanistically, the CoA 3'-phospho ADP moiety interacts with Thr 217, an Aurora A selectivity filter, which permits the formation of an unprecedented covalent bond with Cys 290 in the kinase activation segment, lying some 15 A away. CoA modification (CoAlation) of endogenous Aurora A is rapidly induced by oxidative stresses at Cys 290 in human cells, and microinjection of CoA into mouse embryos perturbs meitoic spindle formation and chromosome alignment. Aurora A regulation by CoA reveals how targeting of Aurora A might be accomplished in the future by development of a 'double-anchored' coval...
The Biochemical journal, Jan 6, 2018
In all living organisms, coenzyme A (CoA) is an essential cofactor with a unique design allowing ... more In all living organisms, coenzyme A (CoA) is an essential cofactor with a unique design allowing it to function as an acyl group carrier and a carbonyl-activating group in diverse biochemical reactions. It is synthesized in a highly conserved process in prokaryotes and eukaryotes that requires pantothenic acid (vitamin B5), cysteine and ATP. CoA and its thioester derivatives are involved in major metabolic pathways, allosteric interactions and the regulation of gene expression. A novel unconventional function of CoA in redox regulation has been recently discovered in mammalian cells and termed protein CoAlation. Here, we report for the first time that protein CoAlation occurs at a background level in exponentially growing bacteria and is strongly induced in response to oxidizing agents and metabolic stress. Over 12% of gene products were shown to be CoAlated in response to diamide-induced stress CoAlation of glyceraldehyde-3-phosphate dehydrogenase was found to inhibit its enzymatic...
PLOS Pathogens
HIV-1 integrates more frequently into transcribed genes, however the biological significance of H... more HIV-1 integrates more frequently into transcribed genes, however the biological significance of HIV-1 integration targeting has remained elusive. Using a selective high-throughput chemical screen, we discovered that the cardiac glycoside digoxin inhibits wild-type HIV-1 infection more potently than HIV-1 bearing a single point mutation (N74D) in the capsid protein. We confirmed that digoxin repressed viral gene expression by targeting the cellular Na + /K + ATPase, but this did not explain its selectivity. Parallel RNAseq and integration mapping in infected cells demonstrated that digoxin inhibited expression of genes involved in Tcell activation and cell metabolism. Analysis of >400,000 unique integration sites showed that WT virus integrated more frequently than N74D mutant within or near genes susceptible to repression by digoxin and involved in T-cell activation and cell metabolism. Two main gene networks down-regulated by the drug were CD40L and CD38. Blocking CD40L by neutralizing antibodies selectively inhibited WT virus infection, phenocopying digoxin. Thus the selectivity of digoxin depends on a combination of integration targeting and repression of specific gene networks. The drug unmasked a functional connection between HIV-1 integration and T-cell activation. Our results suggest that HIV-1 evolved integration site selection to couple its early gene expression with the status of target CD4+ T-cells, which may affect latency and viral reactivation.
The Biochemical journal, Jul 24, 2017
Coenzyme A (CoA) is an obligatory cofactor in all branches of life. CoA and its derivatives are i... more Coenzyme A (CoA) is an obligatory cofactor in all branches of life. CoA and its derivatives are involved in major metabolic pathways, allosteric interactions and the regulation of gene expression. Abnormal biosynthesis and homeostasis of CoA and its derivatives have been associated with various human pathologies, including cancer, diabetes and neurodegeneration. Using an anti-CoA monoclonal antibody and mass spectrometry we identified a wide range of cellular proteins which are modified by covalent attachment of CoA to cysteine thiols (CoAlation). We show that protein CoAlation is a reversible post-translational modification that is induced in mammalian cells and tissues by oxidising agents and metabolic stress. A number of key cellular enzymes were found to be CoAlated in vitro and in vivo in ways that modified their activities. Our study reveals that protein CoAlation is a widespread post-translational modification which may play an important role in redox regulation under physiol...
Retrovirology, 2016
Question: Foamy viruses (FV), and in particular PFV, have emerged in recent years as attractive g... more Question: Foamy viruses (FV), and in particular PFV, have emerged in recent years as attractive gene therapy vector candidates. Since the lack of knowledge on molecular events in FV replication is a major hurdle for broader usage of foamy virus vectors, we aimed at elucidating PFV biology by investigating interactions of its capsid protein, Gag, with host cell components. Methodology and result: To this end, we identified members of the mammalian PLK family as PFV Gag interactants in a commercial yeasttwo-hybrid (Y2H) screen and validated these results in detailed Y2H experiments for PLK1-3. In the yeast system, the intact PLK kinase and substrate recognition motifs were required for interactions with PFV Gag, in which a unique S 224-T-P 226 motif served as a PLK binding determinant. PFV Gag mutants harbouring alanine substitutions of STP residues (iSTP) or phosphomimetic mutations of the T 225 (pmSTP) failed to interact with PLK1-3 in yeast. These findings were corroborated by colocalization studies of ectopically expressed, fluorescently tagged proteins in mammalian cells, where mCherry-tagged PFV Gag was able to recruit eGFP-tagged PLK1 and 2 to condensed mitotic chromatin in an STP motif-dependent manner. When characterizing PFV virions containing wild type or STP mutant Gag proteins, we observed that the mutations did not interfere with particle assembly, release or reverse transcription, but led to a 70 % titer reduction relative to wild type in single-round infection experiments. These replication defects became more prominent in the replication-competent PFV context. Therefore, the lack of Gag STP mutant interaction with PLK proteins upon viral entry into host cells was likely underlying this replication deficit. This hypothesis was strengthened by the finding that enzymatic PLK inhibition in host cells during transduction with wild type PFV mimicked the replication phenotype of PFV STP mutants. In addition to the overall reduced infectivity of the mutants, we also observed that the STP mutations in particle-associated Gag lead to differential sensitivity to integrase inhibition by dolutegravir and resulted in decreased integration efficiency. Conclusions: Taken together, our results demonstrate that PLK proteins influence PFV replication by virtue of their interaction with the Gag protein, ensuring timely and efficient transduction. O2 A novel entry/uncoating assay reveals the presence of at least two species of viral capsids during synchronized HIV-1 infection
Euphytica, 2000
The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan a... more The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan and Taiwan was examined with RAPD-PCR (Random Amplified Polymorphic DNA Polymerase Chain Reaction) markers. Out of the 50 primers screened, 17 primers generated 58 polymorphic and reproducible bands. A minimum of 3 primers was sufficient to distinguish all the 27 accessions studied. The Shannon's index used to partition diversity into inter-and intra-group, revealed that 71 percent of variability resided within groups and 29 percent between groups. Diversity was greatest within the Korean group followed by Taiwan and Japan. The relatively high diversity observed in Korea might reflect the larger genetic base of its plantations while the low diversity in Japan could be explained by the long and intensive tea selection programme in this country. A dendrogram based on the UPGMA-link method using Jaccard's distances and multivariate Factorial correspondence analysis clustered the tea accessions into two main groups, regrouping the Taiwan cultivars on the one side and the Korean and Japanese accessions on the other side. This suggests that the Taiwan tea studied here may have a different origin from that of Korea and Japan.
Molekuliarnaia genetika, mikrobiologiia i virusologiia
Biopolymers and Cell
Розроблено систему оцінки та порівняно in vivo регуляторну здатність рибосомних білків L10. Кліти... more Розроблено систему оцінки та порівняно in vivo регуляторну здатність рибосомних білків L10. Клітини Escherichia coli котрансформуються плазмідами-продуцентами LlO та плазмідою з репортерним геном rplJ'-IacZ. Регуляторна здатність оцінюється за ефективністю пригнічування білком LlO експресії репортерного гену.
Molekuliarnaia genetika, mikrobiologiia i virusologiia
The recombinant plasmids pUC and pNM481 were constructed. The efficiency of P1ac promoter of the ... more The recombinant plasmids pUC and pNM481 were constructed. The efficiency of P1ac promoter of the vector plasmid pUC and PL10. supporting the transcription of rplJL-rpoBC-operons of Escherichia coli was compared. It was found that the stronger promoter P1ac, due to convergent transcription directed by the promoter, makes possible the cloning on the multicopy plasmid pUC of the DNA fragment (the gene rplJ controlled by its own promoter PL10), coding for the synthesis of the regulatory ribosomal protein L10.
Biopolymers and Cell
Короткі повідомлення УДК 677.21 Α. Μ. Живолуп, Є. Б. Патон МОЖЛИВІСТЬ ГЕТЕРОЛОГІЧНОЇ РЕГУЛЯЦІЇ ЕК... more Короткі повідомлення УДК 677.21 Α. Μ. Живолуп, Є. Б. Патон МОЖЛИВІСТЬ ГЕТЕРОЛОГІЧНОЇ РЕГУЛЯЦІЇ ЕКСПРЕСІЇ ГЕНІВ ОПЕРОНУ гр1JL KLEBSIELLA PNEUMONIAE БІЛКОМ L10 ESCHERICHIA COLI Показано можливість досягнення високої ефективності трансформації К. pneumoniae методом електропорації. Виявлено регуляторну здатність білків LlO Е. соїі з нативною та зміненою первинною структурою стосовно генів оперону rplJL К. pneumoniae.
Experimental oncology, 2006
To clone, express in baculovirus expression system and purify SSX2 tumor antigen and to use it fo... more To clone, express in baculovirus expression system and purify SSX2 tumor antigen and to use it for screening of blood serum of melanoma patients. Cloning and expression of SSX2 antigen in Bac-to-Bac baculovirus expression system as His-tag fusion protein, expression of recombinant SSX2 in insect cells with following purification by affinity chromatography on Ni-NTA agarose, ELISA of blood serum of melanoma patients (n = 29) and healthy donors (n = 27) were used. SSX2 was cloned in baculovirus expression system, expressed, purified using affinity chromatography and used in ELISA as antigen. Comparative analysis of blood serum of melanoma patients and healthy donors revealed higher level of SSX2-positivity of blood serum from the patients with cancer. SSX2 antigen expressed in baculovirus expression system can be used for serological analysis of blood serum of oncological patients.
Genetika, 1997
Regulatory interaction of the L10 protein and translation operator of L10 mRNA was studied in viv... more Regulatory interaction of the L10 protein and translation operator of L10 mRNA was studied in vivo using a double-plasmid system. Feedback regulation of rplJ gene expression in Thermotoga maritima was proved, and the possibility of feedback cross-regulation was demonstrated in a heterologous system containing T. maritima and enterobacterial components.