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
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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1978
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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 |
work_keys_str_mv |
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