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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todo:paper_03050491_v127_n2_p155_Sorianello2023-10-03T15:21:14Z Function and structure of rat hepatic coproporphyrinogen oxidase Sorianello, E.M. Mazzetti, M.B. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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, E.M. Mazzetti, M.B. 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. |
| format |
JOUR |
| author |
Sorianello, E.M. Mazzetti, M.B. |
| author_facet |
Sorianello, E.M. Mazzetti, M.B. |
| author_sort |
Sorianello, E.M. |
| 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 |
| url |
http://hdl.handle.net/20.500.12110/paper_03050491_v127_n2_p155_Sorianello |
| work_keys_str_mv |
AT sorianelloem functionandstructureofrathepaticcoproporphyrinogenoxidase AT mazzettimb functionandstructureofrathepaticcoproporphyrinogenoxidase |
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1807319864001429504 |