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Real-time kinetic analysis of macromolecular interaction by using surface plasmon resonance
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Abstracts
activity of the total extract couldn't be determined by Schales' method. The activity assessment of the recombinant proteins needed more optimization. Conclusion: We concluded that although purified natural form of grape class IV chitinase shows an acceptable chitinolytic activity, proper folding of its recombinant forms is crucial for application of this enzyme in biotechnology. Keywords: Grape, Chitinase, Activity, Recombinant, Natural doi:10.1016/j.clinbiochem.2011.08.619
Poster – [A-10-1079-1] Real-time kinetic analysis of macromolecular interaction by using surface plasmon resonance Malihe Keramati 12 Farvardin Ave, Pasteur Institute of Iran, Tehran, Iran E-mail address: [email protected] Bimolecular interaction analysis is at the core of many research projects. The surface-sensitive optical technique of surface plasmon resonance (SPR) is an exciting method for the study of affinity binding interactions. SPR is a phenomenon that occurs in thin conducting films placed at the interface between two media of different refractive indices. In this technique the template is immobilized on the surface, transforming of ligand and template–ligand interaction changes the refractive index that is detected and evaluated by sensorgram curves. Typically, proteinbinding studies make use of fluorescent tags or radiolabels for detection purposes, however SPR-based instrument permit the analysis of biomolecular interaction in real time without labeling, therefore it doesn't alter the protein's active binding site and biological activity and it is possible to mimic the biological environment. This new technology has been widely applied to monitor a diverse set of interaction partners of biological interest, such as protein–protein, protein–lipids, protein– nucleic acids or DNA–DNA, DNA–RNA, protein and low molecular weight molecules such as drugs, substrates and cofactors as well as surface enzyme reactions. In addition SPR biosensors have been used to study the kinetics and affinity of molecular interactions; for identification of novel binding partners by direct capture of proteins from cell or tissue extract on SPR surfaces, hence it is evolving widespread application in ligand fishing, host pathogen interaction, epitope mapping and drug discovery. The increasing number of published papers shows SPR's significant impact in basic research. In this paper the principle and SPR applications will be briefly summarized. Keywords: Biomolecular interaction, Surface plasmon resonance, Kinetic interaction doi:10.1016/j.clinbiochem.2011.08.620
Poster — [A-10-1088-1] Recombinant erythropoietin effects on serum paraoxonase and arylesterase activity, in male rats Jaber Hashemzadeha, Nadereh Rashtchizadeha, Amir Ghorbanihagjoa, Morteza Gharighoranb, Shabnam Hashemzadehc a Medicine Faculty, Tabriz University of Medical Sciences, Tabriz, Iran b Razi University, Iran c Chemistry Faculty, Tabriz University, Iran E-mail address: [email protected] (J. Hashemzadeh) Introduction: Erythropoietin (EPO) is a glycoprotein hormone that is secreted by renal tissue in response to hypoxia and causes
increase in red blood cells (RBC) production. Recent studies demonstrate that EPO has multiple effects such as antiapoptotic, antioxidant, cardioprotective and neuroprotective effects on the body. Paraoxonase (PON), an antioxidant enzyme is secreted by the liver and hydrolyze the lipid peroxidation metabolites. This enzyme has a negative correlation with oxidative stress. The aim of this study is the evaluation of recombinant erythropoietin effects on serum PON and AE activities, in rats. Methods: 30 male adult Wistar rats, were divided into 3 groups, randomly (n = 10). Groups A (high dose r-EPO) and B (low dose rEPO) were injected as intraperitoneal (i.p.) for one and four weeks, respectively. Control group (C) was injected in equal volume normal saline, intraperitoneally. Then, changes in PON and arylesterase (AE) activities, were analyzed. Finally, the differences between three groups were analyzed by SPSS 16.0 softwar. Results: Data analyses showed that, in treated groups with high and low doses of r-EPO in contrast with the control group, there is a significant increase (P < 0.05) in PON–AE activity, statistically also, we observed in this study that in groups A and B that received high and low doses of r-EPO, there was a positive correlation between PON and AE activities, significantly. Conclusion: Based on the results of the current study, recombinant erythropoietin (r-EPO) has antioxidant properties and decrease of lipid peroxidation. Therefore, it can be use in oxidative stress conditions. In spite of expensive cost for use of high dose r-EPO in contrast with low dose, its effects are parallel with low dose r-EPO. This study doesn't suggest use of high dose r-EPO. Keywords: Recombinant erythropoietin (r-EPO), Paraoxonase (PON), Arylesterase (AE) doi:10.1016/j.clinbiochem.2011.08.621
Poster — [A-10-1088-7] Study of relation between serum HDL concentration and paraoxonase activity changes following recombinant erythropoietin injection in male rats Jaber Hashem Zadeha, Nadereh Rashtchizadeha, Amir Ghorbanihagjoa, Morteza Gharighoranb a Biochemistry Department, Tabriz University of Medical Sciences, Tabriz, Iran b Basic Sciences Faculty, Razi University, Iran E-mail address: [email protected] (J. Hashem Zadeh) Introduction: Erythropoietin (EPO) hormone is secreted from renal pretubular tissue and is the main mediator of physiologic response to chronic hypoxia conditions in the body. We studied recombinant erythropoietin effects on serum HDL concentration and paraoxonase(PON) activity in male rats. PON is one of the most significant enzymes in scavenging of harmful free radicals and conserves blood lipoproteins against oxidant components. PON is a HDL-associated enzyme and its activity accounts for antioxidant activity of HDL. The major antioxidant factors associated with high density lipoprotein(HDL). This factors decreased in the serum of patients who have high levels of oxidative stress. Materials and methods: 30 male adult Wistar rats, were divided into 3 groups, randomly (n = 10). Groups A (high dose r-EPO) and B (low dose r-EPO) were injected as intraperitoneal (i.p.) for one and four weeks, respectively. Control group (C) was injected in equal volume normal saline, intraperitoneally. After that, changes in serum PON activity and HDL level were analyzed. Finally, the differences between three groups were analyzed by SPSS 16.0 softwar. Results: Data analyses showed that, in treated groups with high and low doses of r-EPO in contrast with control group, there is a
Abstracts
activity of the total extract couldn't be determined by Schales' method. The activity assessment of the recombinant proteins needed more optimization. Conclusion: We concluded that although purified natural form of grape class IV chitinase shows an acceptable chitinolytic activity, proper folding of its recombinant forms is crucial for application of this enzyme in biotechnology. Keywords: Grape, Chitinase, Activity, Recombinant, Natural doi:10.1016/j.clinbiochem.2011.08.619
Poster – [A-10-1079-1] Real-time kinetic analysis of macromolecular interaction by using surface plasmon resonance Malihe Keramati 12 Farvardin Ave, Pasteur Institute of Iran, Tehran, Iran E-mail address: [email protected] Bimolecular interaction analysis is at the core of many research projects. The surface-sensitive optical technique of surface plasmon resonance (SPR) is an exciting method for the study of affinity binding interactions. SPR is a phenomenon that occurs in thin conducting films placed at the interface between two media of different refractive indices. In this technique the template is immobilized on the surface, transforming of ligand and template–ligand interaction changes the refractive index that is detected and evaluated by sensorgram curves. Typically, proteinbinding studies make use of fluorescent tags or radiolabels for detection purposes, however SPR-based instrument permit the analysis of biomolecular interaction in real time without labeling, therefore it doesn't alter the protein's active binding site and biological activity and it is possible to mimic the biological environment. This new technology has been widely applied to monitor a diverse set of interaction partners of biological interest, such as protein–protein, protein–lipids, protein– nucleic acids or DNA–DNA, DNA–RNA, protein and low molecular weight molecules such as drugs, substrates and cofactors as well as surface enzyme reactions. In addition SPR biosensors have been used to study the kinetics and affinity of molecular interactions; for identification of novel binding partners by direct capture of proteins from cell or tissue extract on SPR surfaces, hence it is evolving widespread application in ligand fishing, host pathogen interaction, epitope mapping and drug discovery. The increasing number of published papers shows SPR's significant impact in basic research. In this paper the principle and SPR applications will be briefly summarized. Keywords: Biomolecular interaction, Surface plasmon resonance, Kinetic interaction doi:10.1016/j.clinbiochem.2011.08.620
Poster — [A-10-1088-1] Recombinant erythropoietin effects on serum paraoxonase and arylesterase activity, in male rats Jaber Hashemzadeha, Nadereh Rashtchizadeha, Amir Ghorbanihagjoa, Morteza Gharighoranb, Shabnam Hashemzadehc a Medicine Faculty, Tabriz University of Medical Sciences, Tabriz, Iran b Razi University, Iran c Chemistry Faculty, Tabriz University, Iran E-mail address: [email protected] (J. Hashemzadeh) Introduction: Erythropoietin (EPO) is a glycoprotein hormone that is secreted by renal tissue in response to hypoxia and causes
increase in red blood cells (RBC) production. Recent studies demonstrate that EPO has multiple effects such as antiapoptotic, antioxidant, cardioprotective and neuroprotective effects on the body. Paraoxonase (PON), an antioxidant enzyme is secreted by the liver and hydrolyze the lipid peroxidation metabolites. This enzyme has a negative correlation with oxidative stress. The aim of this study is the evaluation of recombinant erythropoietin effects on serum PON and AE activities, in rats. Methods: 30 male adult Wistar rats, were divided into 3 groups, randomly (n = 10). Groups A (high dose r-EPO) and B (low dose rEPO) were injected as intraperitoneal (i.p.) for one and four weeks, respectively. Control group (C) was injected in equal volume normal saline, intraperitoneally. Then, changes in PON and arylesterase (AE) activities, were analyzed. Finally, the differences between three groups were analyzed by SPSS 16.0 softwar. Results: Data analyses showed that, in treated groups with high and low doses of r-EPO in contrast with the control group, there is a significant increase (P < 0.05) in PON–AE activity, statistically also, we observed in this study that in groups A and B that received high and low doses of r-EPO, there was a positive correlation between PON and AE activities, significantly. Conclusion: Based on the results of the current study, recombinant erythropoietin (r-EPO) has antioxidant properties and decrease of lipid peroxidation. Therefore, it can be use in oxidative stress conditions. In spite of expensive cost for use of high dose r-EPO in contrast with low dose, its effects are parallel with low dose r-EPO. This study doesn't suggest use of high dose r-EPO. Keywords: Recombinant erythropoietin (r-EPO), Paraoxonase (PON), Arylesterase (AE) doi:10.1016/j.clinbiochem.2011.08.621
Poster — [A-10-1088-7] Study of relation between serum HDL concentration and paraoxonase activity changes following recombinant erythropoietin injection in male rats Jaber Hashem Zadeha, Nadereh Rashtchizadeha, Amir Ghorbanihagjoa, Morteza Gharighoranb a Biochemistry Department, Tabriz University of Medical Sciences, Tabriz, Iran b Basic Sciences Faculty, Razi University, Iran E-mail address: [email protected] (J. Hashem Zadeh) Introduction: Erythropoietin (EPO) hormone is secreted from renal pretubular tissue and is the main mediator of physiologic response to chronic hypoxia conditions in the body. We studied recombinant erythropoietin effects on serum HDL concentration and paraoxonase(PON) activity in male rats. PON is one of the most significant enzymes in scavenging of harmful free radicals and conserves blood lipoproteins against oxidant components. PON is a HDL-associated enzyme and its activity accounts for antioxidant activity of HDL. The major antioxidant factors associated with high density lipoprotein(HDL). This factors decreased in the serum of patients who have high levels of oxidative stress. Materials and methods: 30 male adult Wistar rats, were divided into 3 groups, randomly (n = 10). Groups A (high dose r-EPO) and B (low dose r-EPO) were injected as intraperitoneal (i.p.) for one and four weeks, respectively. Control group (C) was injected in equal volume normal saline, intraperitoneally. After that, changes in serum PON activity and HDL level were analyzed. Finally, the differences between three groups were analyzed by SPSS 16.0 softwar. Results: Data analyses showed that, in treated groups with high and low doses of r-EPO in contrast with control group, there is a