Trypanosoma cruzi adenylate cyclase activity. Purification and characterization

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: sedimen...

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Detalles Bibliográficos
Autores principales:	Torruella, Mónica, Flawiá, Mirtha María, Molina y Vedia, Luis Miguel, Rubinstein, Clara Patricia, Torres, Héctor Norberto
Publicado:	1986
Materias:	adenylate cyclase enzyme monoclonal antibody animal cell chromatography electrophoresis nonhuman priority journal protozoon Trypanosoma cruzi Animalia Eukaryota Protozoa Trypanosoma
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v234_n1_p145_Torruella http://hdl.handle.net/20.500.12110/paper_02646021_v234_n1_p145_Torruella
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_02646021_v234_n1_p145_Torruella
record_format	dspace
spelling	paper:paper_02646021_v234_n1_p145_Torruella2023-06-08T15:23:06Z Trypanosoma cruzi adenylate cyclase activity. Purification and characterization Torruella, Mónica Flawiá, Mirtha María Molina y Vedia, Luis Miguel Rubinstein, Clara Patricia Torres, Héctor Norberto adenylate cyclase enzyme monoclonal antibody animal cell chromatography electrophoresis nonhuman priority journal protozoon Trypanosoma cruzi Animalia Eukaryota Protozoa Trypanosoma Trypanosoma cruzi 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. Fil:Torruella, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Flawia, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Molina y Vedia, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Rubinstein, C.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Torres, H.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1986 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v234_n1_p145_Torruella http://hdl.handle.net/20.500.12110/paper_02646021_v234_n1_p145_Torruella
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	adenylate cyclase enzyme monoclonal antibody animal cell chromatography electrophoresis nonhuman priority journal protozoon Trypanosoma cruzi Animalia Eukaryota Protozoa Trypanosoma Trypanosoma cruzi
spellingShingle	adenylate cyclase enzyme monoclonal antibody animal cell chromatography electrophoresis nonhuman priority journal protozoon Trypanosoma cruzi Animalia Eukaryota Protozoa Trypanosoma Trypanosoma cruzi Torruella, Mónica Flawiá, Mirtha María Molina y Vedia, Luis Miguel Rubinstein, Clara Patricia Torres, Héctor Norberto Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
topic_facet	adenylate cyclase enzyme monoclonal antibody animal cell chromatography electrophoresis nonhuman priority journal protozoon Trypanosoma cruzi Animalia Eukaryota Protozoa Trypanosoma Trypanosoma cruzi
description	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.
author	Torruella, Mónica Flawiá, Mirtha María Molina y Vedia, Luis Miguel Rubinstein, Clara Patricia Torres, Héctor Norberto
author_facet	Torruella, Mónica Flawiá, Mirtha María Molina y Vedia, Luis Miguel Rubinstein, Clara Patricia Torres, Héctor Norberto
author_sort	Torruella, Mónica
title	Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
title_short	Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
title_full	Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
title_fullStr	Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
title_full_unstemmed	Trypanosoma cruzi adenylate cyclase activity. Purification and characterization
title_sort	trypanosoma cruzi adenylate cyclase activity. purification and characterization
publishDate	1986
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v234_n1_p145_Torruella http://hdl.handle.net/20.500.12110/paper_02646021_v234_n1_p145_Torruella
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_version_	1768545463171022848

Trypanosoma cruzi adenylate cyclase activity. Purification and characterization

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