Adenylate cyclase activity associated with Trypanosoma cruzi sedimentable fractions was solubilized by treatment with the non-ionic detergent Lubrol PX and 0.5 M-(NH4)2SO4. The following hydrodynamic and molecular parameters were established for a partially purified enzyme-detergent complex: sedimentation coefficient 6.2 S; Stokes radius 5.65 nm; partial specific volume 0.83 ml/g; M(r) 244000; frictional ratio 1.33. A M(r) of about 124000 was calculated for the detergent-free protein from these parameters. The pI of this enzyme activity was 6.2. A monoclonal antibody to T. cruzi adenylate cyclase was obtained, which inhibited cyclase activities from several lower eukaryotic organisms. The T. cruzi adenylate cyclase was further purified by using this antibody in immunoaffinity chromatographic columns. Fractions obtained after this chromatography showed, on SDS/polyacrylamide-gel electrophoresis, a main polypeptide band with an apparent M(r) of about 56000, which specifically reacted with the monoclonal antibody.
Documento: | Artículo |
Título: | Trypanosoma cruzi adenylate cyclase activity. Purification and characterization |
Autor: | Torruella, M.; Flawia, M.M.; Eisenschlos, C.; Molina y Vedia, L.; Rubinstein, C.P.; Torres, H.N. |
Filiación: | Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, 1428 Buenos Aires, Argentina |
Palabras clave: | adenylate cyclase; enzyme; monoclonal antibody; animal cell; chromatography; electrophoresis; nonhuman; priority journal; protozoon; Trypanosoma cruzi; Animalia; Eukaryota; Protozoa; Trypanosoma; Trypanosoma cruzi |
Año: | 1986 |
Volumen: | 234 |
Número: | 1 |
Página de inicio: | 145 |
Página de fin: | 150 |
DOI: | http://dx.doi.org/10.1042/bj2340145 |
Título revista: | Biochemical Journal |
Título revista abreviado: | BIOCHEM. J. |
ISSN: | 02646021 |
CODEN: | BIJOA |
CAS: | adenylate cyclase, 9012-42-4 |
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v234_n1_p145_Torruella |