Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity

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1. 1. Evidence for dissociation, renaturation, re-association and re-hybridization of bovine liver aminolaevulinate dehydratase attached to Sepharose 4B is reported. 2. 2. When insolubilized enzyme was treated with 3 and 6 M urea, non covalently bound subunits were dissociated and detected in the el...

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Autores principales: Del C. Batlle, A.M., Stella, A.M., Ferramola, A.M., Sopena, Y., De Xifra, E.A.W., Sancovich, H.A.
Formato: Artículo publishedVersion
Lenguaje:Inglés
Publicado: 1978
Materias:
enzyme
porphobilinogen synthase
porphyrin
animal experiment
cattle
liver
Animals
Cattle
Chemistry
Enzymes, Immobilized
Hydrogen-Ion Concentration
Kinetics
Liver
Macromolecular Substances
Models, Chemical
Porphobilinogen Synthase
Protein Denaturation
Sepharose
Solubility
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_0020711X_v9_n6_p401_DelCBatlle
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paperaa:paper_0020711X_v9_n6_p401_DelCBatlle
record_format dspace
spelling paperaa:paper_0020711X_v9_n6_p401_DelCBatlle2023-06-12T16:42:16Z Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity Int. J. Biochem. 1978;9(6):401-406 Del C. Batlle, A.M. Stella, A.M. Ferramola, A.M. Sopena, Y. De Xifra, E.A.W. Sancovich, H.A. enzyme porphobilinogen synthase porphyrin animal experiment cattle liver Animals Cattle Chemistry Enzymes, Immobilized Hydrogen-Ion Concentration Kinetics Liver Macromolecular Substances Models, Chemical Porphobilinogen Synthase Protein Denaturation Sepharose Solubility 1. 1. Evidence for dissociation, renaturation, re-association and re-hybridization of bovine liver aminolaevulinate dehydratase attached to Sepharose 4B is reported. 2. 2. When insolubilized enzyme was treated with 3 and 6 M urea, non covalently bound subunits were dissociated and detected in the eluate; these subunits can be re-associated into a soluble functioning enzyme with a specific activity close to that of the original pure soluble dehydratase preparation. 3. 3. After being washed with a renaturing buffer mixture, the matrix-bound subunits recovered a level of enzymatic activity equal to 50 and 20% of that of the immobilized native aminolaevulinate dehydratase. 4. 4. The reversibility of the dissociation process was investigated. Bound-subunits dehydratase can associate with nascent soluble bovine liver aminolaevulinate dehydratase subunits in situ. The product of such treatment, bound-re-associated enzyme, has the same activity as that of the original bound-dehydratase. The matrix-bound-dissociated bovine liver enzyme was also re-hybridized with soluble dehydratase subunits from E. gracilis. 5. 5. The apparent Km and optimum pH of the immobilized subunits were the same as those of the bound-octameric enzyme. 6. 6. A scheme is proposed, explaining the sequence of reactions leading from the bound-octameric dehydratase to the possible different derivatives, formed during the dissociation and re-association experiments. © 1978. Fil:Del C. Batlle, A.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Stella, A.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ferramola, A.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Sancovich, H.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1978 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0020711X_v9_n6_p401_DelCBatlle
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language Inglés
orig_language_str_mv eng
topic enzyme
porphobilinogen synthase
porphyrin
animal experiment
cattle
liver
Animals
Cattle
Chemistry
Enzymes, Immobilized
Hydrogen-Ion Concentration
Kinetics
Liver
Macromolecular Substances
Models, Chemical
Porphobilinogen Synthase
Protein Denaturation
Sepharose
Solubility
spellingShingle enzyme
porphobilinogen synthase
porphyrin
animal experiment
cattle
liver
Animals
Cattle
Chemistry
Enzymes, Immobilized
Hydrogen-Ion Concentration
Kinetics
Liver
Macromolecular Substances
Models, Chemical
Porphobilinogen Synthase
Protein Denaturation
Sepharose
Solubility
Del C. Batlle, A.M.
Stella, A.M.
Ferramola, A.M.
Sopena, Y.
De Xifra, E.A.W.
Sancovich, H.A.
Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
topic_facet enzyme
porphobilinogen synthase
porphyrin
animal experiment
cattle
liver
Animals
Cattle
Chemistry
Enzymes, Immobilized
Hydrogen-Ion Concentration
Kinetics
Liver
Macromolecular Substances
Models, Chemical
Porphobilinogen Synthase
Protein Denaturation
Sepharose
Solubility
description 1. 1. Evidence for dissociation, renaturation, re-association and re-hybridization of bovine liver aminolaevulinate dehydratase attached to Sepharose 4B is reported. 2. 2. When insolubilized enzyme was treated with 3 and 6 M urea, non covalently bound subunits were dissociated and detected in the eluate; these subunits can be re-associated into a soluble functioning enzyme with a specific activity close to that of the original pure soluble dehydratase preparation. 3. 3. After being washed with a renaturing buffer mixture, the matrix-bound subunits recovered a level of enzymatic activity equal to 50 and 20% of that of the immobilized native aminolaevulinate dehydratase. 4. 4. The reversibility of the dissociation process was investigated. Bound-subunits dehydratase can associate with nascent soluble bovine liver aminolaevulinate dehydratase subunits in situ. The product of such treatment, bound-re-associated enzyme, has the same activity as that of the original bound-dehydratase. The matrix-bound-dissociated bovine liver enzyme was also re-hybridized with soluble dehydratase subunits from E. gracilis. 5. 5. The apparent Km and optimum pH of the immobilized subunits were the same as those of the bound-octameric enzyme. 6. 6. A scheme is proposed, explaining the sequence of reactions leading from the bound-octameric dehydratase to the possible different derivatives, formed during the dissociation and re-association experiments. © 1978.
format Artículo
Artículo
publishedVersion
author Del C. Batlle, A.M.
Stella, A.M.
Ferramola, A.M.
Sopena, Y.
De Xifra, E.A.W.
Sancovich, H.A.
author_facet Del C. Batlle, A.M.
Stella, A.M.
Ferramola, A.M.
Sopena, Y.
De Xifra, E.A.W.
Sancovich, H.A.
author_sort Del C. Batlle, A.M.
title Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
title_short Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
title_full Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
title_fullStr Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
title_full_unstemmed Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
title_sort porphyrin biosynthesis-immobilized enzymes and ligands-x. a novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity
publishDate 1978
url http://hdl.handle.net/20.500.12110/paper_0020711X_v9_n6_p401_DelCBatlle
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AT stellaam porphyrinbiosynthesisimmobilizedenzymesandligandsxanovelapproachtothestudyoftherelationshipbetweenthequaternarystructureofaminolaevulinatedehydrataseanditsactivity
AT ferramolaam porphyrinbiosynthesisimmobilizedenzymesandligandsxanovelapproachtothestudyoftherelationshipbetweenthequaternarystructureofaminolaevulinatedehydrataseanditsactivity
AT sopenay porphyrinbiosynthesisimmobilizedenzymesandligandsxanovelapproachtothestudyoftherelationshipbetweenthequaternarystructureofaminolaevulinatedehydrataseanditsactivity
AT dexifraeaw porphyrinbiosynthesisimmobilizedenzymesandligandsxanovelapproachtothestudyoftherelationshipbetweenthequaternarystructureofaminolaevulinatedehydrataseanditsactivity
AT sancovichha porphyrinbiosynthesisimmobilizedenzymesandligandsxanovelapproachtothestudyoftherelationshipbetweenthequaternarystructureofaminolaevulinatedehydrataseanditsactivity
_version_ 1769810012387934208

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