Function and structure of rat hepatic coproporphyrinogen oxidase

Rat hepatic coproporphyrinogen oxidase, the sixth enzyme in the heme biosynthetic pathway, was purified 1340-fold with a yield of 39.7%. To obtain the soluble enzyme, different methods were applied to disrupt mitochondria, with sonication giving the highest yield (85%). The minimum catalytic form of...

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Autores principales: Sorianello, Eleonora Mariana, Mazzetti, Marta Blanca
Publicado: 2000
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rat
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03050491_v127_n2_p155_Sorianello
http://hdl.handle.net/20.500.12110/paper_03050491_v127_n2_p155_Sorianello
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spelling paper:paper_03050491_v127_n2_p155_Sorianello2023-06-08T15:30:18Z Function and structure of rat hepatic coproporphyrinogen oxidase Sorianello, Eleonora Mariana Mazzetti, Marta Blanca Amino acid modifiers Catalytic activity Coproporphyria Coproporphyrin Coproporphyrinogen oxidase Enzyme Protoporphyrin IX Protoporphyrinogen IX Purification Rat liver coproporphyrin coproporphyrinogen oxidase protoporphyrinogen oxidase amino acid sequence animal experiment animal tissue article controlled study coproporphyria enzyme activity enzyme purification enzyme structure, function and variability liver metabolism molecular weight nonhuman priority journal rat Animals Chromatography, High Pressure Liquid Coproporphyrinogen Oxidase Detergents Diethyl Pyrocarbonate Humans Liver Octoxynol Phenylglyoxal Phospholipids Polysorbates Rats Rats, Sprague-Dawley Water Rat hepatic coproporphyrinogen oxidase, the sixth enzyme in the heme biosynthetic pathway, was purified 1340-fold with a yield of 39.7%. To obtain the soluble enzyme, different methods were applied to disrupt mitochondria, with sonication giving the highest yield (85%). The minimum catalytic form of enzyme was a dimer with a molecular mass of 77±4 kDa. The existence of aggregated forms was possible since in fractions of gel filtration elution activity was observed with higher molecular mass. We determined a Stokes radius of 36.3 A, a sedimentation coefficient (S20,(w)) of 5.06 S, and frictional ratio of 1.29, suggesting a nearly globular shape of the protein. Regardless of the type of salt, high ionic strength inhibits the enzyme, probably modifying its native structure. Experiments with amino acid modifiers showed that histidine, arginine, and tryptophan are involved in the catalytic process. Non-ionic detergents and phospholipids activated the enzyme, probably because they reproduce its natural hydrophobic environment. The present study describes a simple method for the purification of rat liver coproporphyrinogen oxidase, introducing for the first time data on the structure and function of the protein in a tissue often used as a laboratory model in biological studies, and contributing to the study of human hereditary coproporphyria. Copyright (C) 2000 Elsevier Science Inc. Fil:Sorianello, E.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Mazzetti, M.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2000 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03050491_v127_n2_p155_Sorianello http://hdl.handle.net/20.500.12110/paper_03050491_v127_n2_p155_Sorianello
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Amino acid modifiers
Catalytic activity
Coproporphyria
Coproporphyrin
Coproporphyrinogen oxidase
Enzyme
Protoporphyrin IX
Protoporphyrinogen IX
Purification
Rat liver
coproporphyrin
coproporphyrinogen oxidase
protoporphyrinogen oxidase
amino acid sequence
animal experiment
animal tissue
article
controlled study
coproporphyria
enzyme activity
enzyme purification
enzyme structure, function and variability
liver metabolism
molecular weight
nonhuman
priority journal
rat
Animals
Chromatography, High Pressure Liquid
Coproporphyrinogen Oxidase
Detergents
Diethyl Pyrocarbonate
Humans
Liver
Octoxynol
Phenylglyoxal
Phospholipids
Polysorbates
Rats
Rats, Sprague-Dawley
Water
spellingShingle Amino acid modifiers
Catalytic activity
Coproporphyria
Coproporphyrin
Coproporphyrinogen oxidase
Enzyme
Protoporphyrin IX
Protoporphyrinogen IX
Purification
Rat liver
coproporphyrin
coproporphyrinogen oxidase
protoporphyrinogen oxidase
amino acid sequence
animal experiment
animal tissue
article
controlled study
coproporphyria
enzyme activity
enzyme purification
enzyme structure, function and variability
liver metabolism
molecular weight
nonhuman
priority journal
rat
Animals
Chromatography, High Pressure Liquid
Coproporphyrinogen Oxidase
Detergents
Diethyl Pyrocarbonate
Humans
Liver
Octoxynol
Phenylglyoxal
Phospholipids
Polysorbates
Rats
Rats, Sprague-Dawley
Water
Sorianello, Eleonora Mariana
Mazzetti, Marta Blanca
Function and structure of rat hepatic coproporphyrinogen oxidase
topic_facet Amino acid modifiers
Catalytic activity
Coproporphyria
Coproporphyrin
Coproporphyrinogen oxidase
Enzyme
Protoporphyrin IX
Protoporphyrinogen IX
Purification
Rat liver
coproporphyrin
coproporphyrinogen oxidase
protoporphyrinogen oxidase
amino acid sequence
animal experiment
animal tissue
article
controlled study
coproporphyria
enzyme activity
enzyme purification
enzyme structure, function and variability
liver metabolism
molecular weight
nonhuman
priority journal
rat
Animals
Chromatography, High Pressure Liquid
Coproporphyrinogen Oxidase
Detergents
Diethyl Pyrocarbonate
Humans
Liver
Octoxynol
Phenylglyoxal
Phospholipids
Polysorbates
Rats
Rats, Sprague-Dawley
Water
description Rat hepatic coproporphyrinogen oxidase, the sixth enzyme in the heme biosynthetic pathway, was purified 1340-fold with a yield of 39.7%. To obtain the soluble enzyme, different methods were applied to disrupt mitochondria, with sonication giving the highest yield (85%). The minimum catalytic form of enzyme was a dimer with a molecular mass of 77±4 kDa. The existence of aggregated forms was possible since in fractions of gel filtration elution activity was observed with higher molecular mass. We determined a Stokes radius of 36.3 A, a sedimentation coefficient (S20,(w)) of 5.06 S, and frictional ratio of 1.29, suggesting a nearly globular shape of the protein. Regardless of the type of salt, high ionic strength inhibits the enzyme, probably modifying its native structure. Experiments with amino acid modifiers showed that histidine, arginine, and tryptophan are involved in the catalytic process. Non-ionic detergents and phospholipids activated the enzyme, probably because they reproduce its natural hydrophobic environment. The present study describes a simple method for the purification of rat liver coproporphyrinogen oxidase, introducing for the first time data on the structure and function of the protein in a tissue often used as a laboratory model in biological studies, and contributing to the study of human hereditary coproporphyria. Copyright (C) 2000 Elsevier Science Inc.
author Sorianello, Eleonora Mariana
Mazzetti, Marta Blanca
author_facet Sorianello, Eleonora Mariana
Mazzetti, Marta Blanca
author_sort Sorianello, Eleonora Mariana
title Function and structure of rat hepatic coproporphyrinogen oxidase
title_short Function and structure of rat hepatic coproporphyrinogen oxidase
title_full Function and structure of rat hepatic coproporphyrinogen oxidase
title_fullStr Function and structure of rat hepatic coproporphyrinogen oxidase
title_full_unstemmed Function and structure of rat hepatic coproporphyrinogen oxidase
title_sort function and structure of rat hepatic coproporphyrinogen oxidase
publishDate 2000
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03050491_v127_n2_p155_Sorianello
http://hdl.handle.net/20.500.12110/paper_03050491_v127_n2_p155_Sorianello
work_keys_str_mv AT sorianelloeleonoramariana functionandstructureofrathepaticcoproporphyrinogenoxidase
AT mazzettimartablanca functionandstructureofrathepaticcoproporphyrinogenoxidase
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