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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Autores principales: Torruella, M., Flawia, M.M., Eisenschlos, C., Molina y Vedia, L., Rubinstein, C.P., Torres, H.N.
Formato: JOUR
Materias:
adenylate cyclase
enzyme
monoclonal antibody
animal cell
chromatography
electrophoresis
nonhuman
priority journal
protozoon
Trypanosoma cruzi
Animalia
Eukaryota
Protozoa
Trypanosoma
Acceso en línea: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 todo:paper_02646021_v234_n1_p145_Torruella
record_format dspace
spelling todo:paper_02646021_v234_n1_p145_Torruella2023-10-03T15:12:53Z Trypanosoma cruzi adenylate cyclase activity. Purification and characterization Torruella, M. Flawia, M.M. Eisenschlos, C. Molina y Vedia, L. Rubinstein, C.P. Torres, H.N. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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, M.
Flawia, M.M.
Eisenschlos, C.
Molina y Vedia, L.
Rubinstein, C.P.
Torres, H.N.
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.
format JOUR
author Torruella, M.
Flawia, M.M.
Eisenschlos, C.
Molina y Vedia, L.
Rubinstein, C.P.
Torres, H.N.
author_facet Torruella, M.
Flawia, M.M.
Eisenschlos, C.
Molina y Vedia, L.
Rubinstein, C.P.
Torres, H.N.
author_sort Torruella, M.
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
url http://hdl.handle.net/20.500.12110/paper_02646021_v234_n1_p145_Torruella
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AT eisenschlosc trypanosomacruziadenylatecyclaseactivitypurificationandcharacterization
AT molinayvedial trypanosomacruziadenylatecyclaseactivitypurificationandcharacterization
AT rubinsteincp trypanosomacruziadenylatecyclaseactivitypurificationandcharacterization
AT torreshn trypanosomacruziadenylatecyclaseactivitypurificationandcharacterization
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