Effect of binase on phorbol myristate acetate-induced apoptosis of human peripheral blood granulocytes and monocytes

Effect of binase (RNAse of Bacillus intermedius) on phorbol myristate acetate-(PMA)-induced apoptosis of human peripheral blood granulocytes and monocytes was studied in vitro by flow cytometry. Both toxic (400 μg/ml) and nontoxic (40 μg/ml) binase concentrations were tested. The binase end-point effect was dependent on the target cell population and the binase concentration. In a granulocyte subset, the 400 μg/ml concentration resulted in strongly pronounced stimulation of PMA-induced apoptosis. In a monocyte subset, the 40 μg/ml concentration developed a protective effect as judged by an increase in a percantage of viable cell subset and by slowing-down cells transtion from an early to late PMA-induced apoptotic phase

A hydrophobic segment of some cytotoxic ribonucleases

The exact mechanism by which cytotoxic ribonucleases reach the cytosol of tumor cells remains unclear. The interaction of ribonucleases with a lipid bilayer is involved in the translocation of ribonucleases across the endosomal membrane. Here, we aimed to study the hydropathy character of toxic antitumor ribonucleases (bovine seminal ribonuclease and binase) and two non-toxic ribonucleases (bovine pancreatic ribonuclease and human pancreatic ribonuclease) by sliding-window hydrophobicity analysis. Comparative hydropathy plot analysis of the non-toxic pancreatic ribonucleases and their toxic variants was also performed. The data obtained indicate that some cytotoxic ribonucleases have a hydrophobic segment, which is sterically available for the hydrophobic interaction with a tumor cell membrane and endosomal membrane. After dissociation, subunits of dimeric ribonucleases are probably capable of thermodynamically favorable interaction with the interfacial region of a lipid bilayer. Remarkably the hydrophobic segment is not identified in the amino acid sequences of non-toxic ribonucleases. The paper describes the hydrophobic properties of toxic RNases that are essential for both the model of a lipid-protein interaction and the cytotoxicity mechanism unraveling. © 2013 Elsevier Ltd

Study of characteristics of electrostatic interaction between RNases and mica surface using atomic force microscopy

The physical adsorption of pancreatic RNase A and Bacillus intermedius RNase on a negatively charged mica surface was investigated using atomic force microscopy. An analysis of the kinetics of RNase adsorption showed that Bacillus intermedius RNase was adsorbed 3-5 times more effectively than RNase A. At the same time, the adsorption of Bacillus intermedius RNase on mica was accompanied by the aggregation of enzyme molecules. © 2011 Pleiades Publishing, Ltd

Effect of binase on phorbol myristate acetate-induced apoptosis of human peripheral blood granulocytes and monocytes

Effect of binase (RNAse of Bacillus intermedius) on phorbol myristate acetate-(PMA)-induced apoptosis of human peripheral blood granulocytes and monocytes was studied in vitro by flow cytometry. Both toxic (400 μg/ml) and nontoxic (40 μg/ml) binase concentrations were tested. The binase end-point effect was dependent on the target cell population and the binase concentration. In a granulocyte subset, the 400 μg/ml concentration resulted in strongly pronounced stimulation of PMA-induced apoptosis. In a monocyte subset, the 40 μg/ml concentration developed a protective effect as judged by an increase in a percantage of viable cell subset and by slowing-down cells transtion from an early to late PMA-induced apoptotic phase

Effect of binase on phorbol myristate acetate-induced apoptosis of human peripheral blood granulocytes and monocytes

Effect of binase (RNAse of Bacillus intermedius) on phorbol myristate acetate-(PMA)-induced apoptosis of human peripheral blood granulocytes and monocytes was studied in vitro by flow cytometry. Both toxic (400 μg/ml) and nontoxic (40 μg/ml) binase concentrations were tested. The binase end-point effect was dependent on the target cell population and the binase concentration. In a granulocyte subset, the 400 μg/ml concentration resulted in strongly pronounced stimulation of PMA-induced apoptosis. In a monocyte subset, the 40 μg/ml concentration developed a protective effect as judged by an increase in a percantage of viable cell subset and by slowing-down cells transtion from an early to late PMA-induced apoptotic phase

A hydrophobic segment of some cytotoxic ribonucleases

The exact mechanism by which cytotoxic ribonucleases reach the cytosol of tumor cells remains unclear. The interaction of ribonucleases with a lipid bilayer is involved in the translocation of ribonucleases across the endosomal membrane. Here, we aimed to study the hydropathy character of toxic antitumor ribonucleases (bovine seminal ribonuclease and binase) and two non-toxic ribonucleases (bovine pancreatic ribonuclease and human pancreatic ribonuclease) by sliding-window hydrophobicity analysis. Comparative hydropathy plot analysis of the non-toxic pancreatic ribonucleases and their toxic variants was also performed. The data obtained indicate that some cytotoxic ribonucleases have a hydrophobic segment, which is sterically available for the hydrophobic interaction with a tumor cell membrane and endosomal membrane. After dissociation, subunits of dimeric ribonucleases are probably capable of thermodynamically favorable interaction with the interfacial region of a lipid bilayer. Remarkably the hydrophobic segment is not identified in the amino acid sequences of non-toxic ribonucleases. The paper describes the hydrophobic properties of toxic RNases that are essential for both the model of a lipid-protein interaction and the cytotoxicity mechanism unraveling. © 2013 Elsevier Ltd

Study of characteristics of electrostatic interaction between RNases and mica surface using atomic force microscopy

The physical adsorption of pancreatic RNase A and Bacillus intermedius RNase on a negatively charged mica surface was investigated using atomic force microscopy. An analysis of the kinetics of RNase adsorption showed that Bacillus intermedius RNase was adsorbed 3-5 times more effectively than RNase A. At the same time, the adsorption of Bacillus intermedius RNase on mica was accompanied by the aggregation of enzyme molecules. © 2011 Pleiades Publishing, Ltd

Study of characteristics of electrostatic interaction between RNases and mica surface using atomic force microscopy

The physical adsorption of pancreatic RNase A and Bacillus intermedius RNase on a negatively charged mica surface was investigated using atomic force microscopy. An analysis of the kinetics of RNase adsorption showed that Bacillus intermedius RNase was adsorbed 3-5 times more effectively than RNase A. At the same time, the adsorption of Bacillus intermedius RNase on mica was accompanied by the aggregation of enzyme molecules. © 2011 Pleiades Publishing, Ltd

Study of characteristics of electrostatic interaction between RNases and mica surface using atomic force microscopy

The physical adsorption of pancreatic RNase A and Bacillus intermedius RNase on a negatively charged mica surface was investigated using atomic force microscopy. An analysis of the kinetics of RNase adsorption showed that Bacillus intermedius RNase was adsorbed 3-5 times more effectively than RNase A. At the same time, the adsorption of Bacillus intermedius RNase on mica was accompanied by the aggregation of enzyme molecules. © 2011 Pleiades Publishing, Ltd

A hydrophobic segment of some cytotoxic ribonucleases

The exact mechanism by which cytotoxic ribonucleases reach the cytosol of tumor cells remains unclear. The interaction of ribonucleases with a lipid bilayer is involved in the translocation of ribonucleases across the endosomal membrane. Here, we aimed to study the hydropathy character of toxic antitumor ribonucleases (bovine seminal ribonuclease and binase) and two non-toxic ribonucleases (bovine pancreatic ribonuclease and human pancreatic ribonuclease) by sliding-window hydrophobicity analysis. Comparative hydropathy plot analysis of the non-toxic pancreatic ribonucleases and their toxic variants was also performed. The data obtained indicate that some cytotoxic ribonucleases have a hydrophobic segment, which is sterically available for the hydrophobic interaction with a tumor cell membrane and endosomal membrane. After dissociation, subunits of dimeric ribonucleases are probably capable of thermodynamically favorable interaction with the interfacial region of a lipid bilayer. Remarkably the hydrophobic segment is not identified in the amino acid sequences of non-toxic ribonucleases. The paper describes the hydrophobic properties of toxic RNases that are essential for both the model of a lipid-protein interaction and the cytotoxicity mechanism unraveling. © 2013 Elsevier Ltd