RESUMO
Sequence-specificity of binding of ribonuclease binase to oligodeoxyribonucleotides immobilized in biochip gel pads was studied. Binding constants for the complexes between binase and selected oligonucleotides were measured. Oligodeoxyribonucleotides GAGAGAG and GAGAGAGAG were found to be high-specific in interaction with binase. These oligonucleotides were used as molecular probes for immobilization in a sorptive medium for subsequent isolation and concentrating of binase from diluted water solutions. Volume capacity of the developed sorptive mediums containing immobilized oligodeoxyribonucleotides GAGAGAG and GAGAGAGAG was found to be 2.6 and 2.3 mg of binase per 1 mL of sorbate correspondingly. After the procedure of affinity chromatography and elution ofbinase from sorptive medium the protein showed the same affinity of binding to oligonucleotides as the initial sample.
Assuntos
Proteínas de Ligação a DNA/isolamento & purificação , Endorribonucleases/isolamento & purificação , Análise Serial de Proteínas/métodos , Adsorção , Sequência de Aminoácidos/genética , Animais , Sequência de Bases/genética , Sítios de Ligação/genética , Cromatografia de Afinidade , DNA/metabolismo , Proteínas de Ligação a DNA/farmacocinética , Endorribonucleases/farmacocinética , Humanos , Ácidos Nucleicos Imobilizados/genética , Ácidos Nucleicos Imobilizados/metabolismo , Sondas Moleculares/genética , Ligação ProteicaAssuntos
Anticorpos Biespecíficos , Colágeno/química , Enzimas Imobilizadas/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Ativador de Plasminogênio Tipo Uroquinase/farmacologia , Animais , Enzimas Imobilizadas/química , Humanos , Técnicas In Vitro , Ativação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/química , Ratos , Ativador de Plasminogênio Tipo Uroquinase/químicaRESUMO
A digitonin-containing affinity sorbent was prepared using a hydrophilic polymer carrier. The correlation was found between the polymer structure and crosslink. King degree, on the one hand, and the sorbent capacity to bind cholesterol, protein and lipoproteins--on the other. The sorbent was demonstrated to extract predominantly atherogenic lipoproteins from blood plasma.
Assuntos
Arteriosclerose/sangue , LDL-Colesterol/isolamento & purificação , Colesterol/isolamento & purificação , Hemoperfusão , Lipoproteínas VLDL/isolamento & purificação , Animais , Apolipoproteínas/sangue , Apolipoproteínas/isolamento & purificação , Colesterol/sangue , LDL-Colesterol/sangue , VLDL-Colesterol , Digitonina , Humanos , Hipercolesterolemia/sangue , Lipoproteínas VLDL/sangue , Anidridos Maleicos , Polímeros/análise , Polímeros/síntese química , Pirrolidinonas , CoelhosRESUMO
Industrial silicates, silochromes, proved to be effective sorbents of cholesterol lipoprotein complexes in blood in vitro. Silochrome SH-1.5 with a diameter of pores of 180-200 nm and a capacity of 12-14 mg of cholesterol per 1 g of the sorbent exhibited the best efficiency in isolation of the complexes. At the same time, maximal unspecific binding of blood plasma proteins was observed in the systems with silochrome SH-3 with a diameter of pores of 60 nm. By use of silochrome SH-1.5 for plasmosorption the content of cholesterol was decreased from 3,600 mg/l down to 2,150 mg/l, i.e. by 40%, in blood plasma of a rabbit with experimental hypercholesterolemia.
Assuntos
Colesterol/sangue , Hemoperfusão/métodos , Lipoproteínas/sangue , Dióxido de Silício , Animais , Colesterol/isolamento & purificação , LDL-Colesterol/sangue , LDL-Colesterol/isolamento & purificação , Humanos , Lipoproteínas/isolamento & purificação , Coelhos , Sílica GelRESUMO
The hormones of thyroid gland thyroxine and triiodothyronine were shown to increase the permeability of bilayer lipid membrane (BLM) and inner mitochondrial membrane for hydrophobic positively charged complex K+-nonactine and to decrease the permeability of these membranes for hydrophobic anion FCCP-. These facts imply that the thyroid hormones affect the phospholipid membranes like the dipole modifyers decreasing the positive potential of hydrophobic region of the membranes in respect to the water phase.