RESUMO
The detection of biomarkers to be applied to reveal different types of tumors is an actual demand of today. Since course of this disease can be asymptomatic these kinds of markers can be applied in widely-distributed screening procedures. Many types of biomarkers for detection of cancer exist. However all these markers are specific only for particular type of tumor and have no use in screening procedures for detection of this pathology. The Annexin 5, a Ca-depended phospholipid binding protein, was discovered in serums of all pregnant women on various stages of pregnancy. The study was implemented using method of latex agglutination. More than that, this protein was detected in serums of patients with several types of cancer. The possibility to apply annexin 5 as marker for screening of various types of cancer is considered.
Assuntos
Anexina A5/sangue , Neoplasias/sangue , Gravidez/sangue , Adulto , Feminino , Humanos , Testes de Fixação do Látex , Neoplasias/diagnósticoRESUMO
A method for preparing polystyrene latexes for slide agglutination test is described. The latexes were used for preparing latex slide tests for evaluating C-reactive protein and myoglobin. Effects of such factors as pH, temperature, antibody concentration, and latex particles concentration, on sensitization and agglutination have been studied. The proposed approaches allow detection of the antigen in a concentration of less than 200 micrograms/liter for myoglobin and less than 8 mg/liter for C-reactive protein.
Assuntos
Proteína C-Reativa/análise , Mioglobina/análise , Poliestirenos , Animais , Humanos , Testes de Fixação do Látex , Poliestirenos/síntese química , Coelhos , RatosRESUMO
Some properties of flavin-containing polyamine oxidase from bovine liver have been studied. Limited proteolysis of the enzyme by trypsin resulted in two fragments with molecular masses of 48 and 5-8 kDa. The antigenic determinants of the protein appear to be bound to the larger proteolytic fragment. The effect of proteolysis on optical properties of the enzyme has been established. Data from immunological analysis suggest that the soluble fraction of the liver is a rich source of the enzyme.
Assuntos
Flavinas/metabolismo , Fígado/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Animais , Bovinos , Epitopos/metabolismo , Hidrólise , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/imunologia , Coelhos , Espectrometria de Fluorescência , Tripsina/metabolismo , Poliamina OxidaseRESUMO
A flavin-containing polyamine oxidase was isolated in an electrophoretically homogeneous state from cattle liver cytosol. The molecular mass, subunit composition and flavin content of the enzyme were determined; flavin, was shown to be covalently bound to the protein fragment of the polyamine oxidase molecule. Some optical and luminescent properties of the native and denatured enzyme were investigated. Denaturation and quenching were found to affect the luminescent properties of polyamine oxidase.
Assuntos
Fígado/enzimologia , Proteínas Luminescentes/isolamento & purificação , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/isolamento & purificação , Animais , Bovinos , Flavinas/análise , Proteínas Luminescentes/química , Peso Molecular , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/química , Desnaturação Proteica/fisiologia , Espectrofotometria , Poliamina OxidaseRESUMO
It was demonstrated that the NADPH-adrenodoxin reductase molecule contains ten tryptophan residues titrated by N-bromosuccinimide. The effectiveness of the non-radiant energy transfer was used to calculate the average distance between the NADPH-binding site of the enzyme and tryptophan residues at different steps of N-bromosuccinimide-induced modification.
Assuntos
Ferredoxina-NADP Redutase/análise , NADH NADPH Oxirredutases/análise , Triptofano/análise , Córtex Suprarrenal/enzimologia , Animais , Sítios de Ligação , Bromosuccinimida , Mitocôndrias/enzimologia , NADP , Espectrometria de FluorescênciaRESUMO
The fluorescent and phosphorescent properties of NADPH-adrenodoxin reductase were investigated. It was shown that the fluorescence of protein tryptophanyls was quenched completely by acrylamide and partially by ionic quenchers (I- and Cs+). A removal of the prosthetic group from the protein causes insignificant changes in fluorescent properties of the enzyme. The denaturation of the enzyme by urea was accompanied by growth of quenching parameters. Indeed, some differences were observed in the quenching of flavin fluorescence by ionic quenchers (I- and Cs+). NADPH appeared to be an efficient quencher of NADPH-adrenodoxin reductase tryptophan fluorescence. Using Förster's equations for non-radiative energy transfer, the distance between NADPH-binding site and tryptophanyls was evaluated to 35-40 A.