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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Veröffentlicht in: | Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2000-10, Vol.127 (2), p.155-164 |
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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 Å, a sedimentation coefficient (S
20,
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. |
doi_str_mv | 10.1016/S0305-0491(00)00247-9 |
format | Article |
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20,
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.</description><identifier>ISSN: 1096-4959</identifier><identifier>EISSN: 1879-1107</identifier><identifier>DOI: 10.1016/S0305-0491(00)00247-9</identifier><identifier>PMID: 11079369</identifier><language>eng</language><publisher>England: Elsevier Inc</publisher><subject>Amino acid modifiers ; Animals ; Catalytic activity ; Chromatography, High Pressure Liquid ; Coproporphyria ; Coproporphyrin ; Coproporphyrinogen oxidase ; Coproporphyrinogen Oxidase - chemistry ; Coproporphyrinogen Oxidase - metabolism ; Coproporphyrinogen Oxidase - physiology ; Detergents - pharmacology ; Diethyl Pyrocarbonate - metabolism ; Enzyme ; Humans ; Liver - enzymology ; Octoxynol - pharmacology ; Phenylglyoxal - pharmacology ; Phospholipids - metabolism ; Polysorbates - pharmacology ; Protoporphyrin IX ; Protoporphyrinogen IX ; Purification ; Rat liver ; Rats ; Rats, Sprague-Dawley ; Water - metabolism</subject><ispartof>Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2000-10, Vol.127 (2), p.155-164</ispartof><rights>2000 Elsevier Science Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c361t-f5ef5644f2d6ee64a43b0a8b6582b070849db135a07ed42521161885998000563</citedby><cites>FETCH-LOGICAL-c361t-f5ef5644f2d6ee64a43b0a8b6582b070849db135a07ed42521161885998000563</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0305-0491(00)00247-9$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,782,786,3552,27931,27932,46002</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11079369$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sorianello, Eleonora M</creatorcontrib><creatorcontrib>Mazzetti, Marta B</creatorcontrib><title>Function and structure of rat hepatic coproporphyrinogen oxidase</title><title>Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology</title><addtitle>Comp Biochem Physiol B Biochem Mol Biol</addtitle><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 Å, a sedimentation coefficient (S
20,
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.</description><subject>Amino acid modifiers</subject><subject>Animals</subject><subject>Catalytic activity</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Coproporphyria</subject><subject>Coproporphyrin</subject><subject>Coproporphyrinogen oxidase</subject><subject>Coproporphyrinogen Oxidase - chemistry</subject><subject>Coproporphyrinogen Oxidase - metabolism</subject><subject>Coproporphyrinogen Oxidase - physiology</subject><subject>Detergents - pharmacology</subject><subject>Diethyl Pyrocarbonate - metabolism</subject><subject>Enzyme</subject><subject>Humans</subject><subject>Liver - enzymology</subject><subject>Octoxynol - pharmacology</subject><subject>Phenylglyoxal - pharmacology</subject><subject>Phospholipids - metabolism</subject><subject>Polysorbates - pharmacology</subject><subject>Protoporphyrin IX</subject><subject>Protoporphyrinogen IX</subject><subject>Purification</subject><subject>Rat liver</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Water - metabolism</subject><issn>1096-4959</issn><issn>1879-1107</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EolD4BFBWCBaBcfxIvAJUUUCqxAJYW44zoUZtHOwE0b8nfUgsWc1Ic-_cmUPIGYVrClTevAIDkQJX9BLgCiDjear2yBEtcpVSCvn-0IOSKVdCjchxjJ8ArKCMHpLReq6YVEfkbto3tnO-SUxTJbELve36gImvk2C6ZI6t6ZxNrG-Db31o56vgGv-BTeJ_XGUinpCD2iwinu7qmLxPH94mT-ns5fF5cj9LLZO0S2uBtZCc11klESU3nJVgilKKIishh4KrqqRMGMix4pnIKJW0KIRSBQAIycbkYrt3OOSrx9jppYsWFwvToO-jzjNOOcvYIBRboQ0-xoC1boNbmrDSFPQand6g02t0GkBv0Gk1-M53AX25xOrPtWM1CG63Ahze_HYYdLQOG4uVC2g7XXn3T8Qv6hd8yw</recordid><startdate>20001001</startdate><enddate>20001001</enddate><creator>Sorianello, Eleonora M</creator><creator>Mazzetti, Marta B</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20001001</creationdate><title>Function and structure of rat hepatic coproporphyrinogen oxidase</title><author>Sorianello, Eleonora M ; Mazzetti, Marta B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c361t-f5ef5644f2d6ee64a43b0a8b6582b070849db135a07ed42521161885998000563</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Amino acid modifiers</topic><topic>Animals</topic><topic>Catalytic activity</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Coproporphyria</topic><topic>Coproporphyrin</topic><topic>Coproporphyrinogen oxidase</topic><topic>Coproporphyrinogen Oxidase - chemistry</topic><topic>Coproporphyrinogen Oxidase - metabolism</topic><topic>Coproporphyrinogen Oxidase - physiology</topic><topic>Detergents - pharmacology</topic><topic>Diethyl Pyrocarbonate - metabolism</topic><topic>Enzyme</topic><topic>Humans</topic><topic>Liver - enzymology</topic><topic>Octoxynol - pharmacology</topic><topic>Phenylglyoxal - pharmacology</topic><topic>Phospholipids - metabolism</topic><topic>Polysorbates - pharmacology</topic><topic>Protoporphyrin IX</topic><topic>Protoporphyrinogen IX</topic><topic>Purification</topic><topic>Rat liver</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Water - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sorianello, Eleonora M</creatorcontrib><creatorcontrib>Mazzetti, Marta B</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sorianello, Eleonora M</au><au>Mazzetti, Marta B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Function and structure of rat hepatic coproporphyrinogen oxidase</atitle><jtitle>Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology</jtitle><addtitle>Comp Biochem Physiol B Biochem Mol Biol</addtitle><date>2000-10-01</date><risdate>2000</risdate><volume>127</volume><issue>2</issue><spage>155</spage><epage>164</epage><pages>155-164</pages><issn>1096-4959</issn><eissn>1879-1107</eissn><abstract>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 Å, a sedimentation coefficient (S
20,
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.</abstract><cop>England</cop><pub>Elsevier Inc</pub><pmid>11079369</pmid><doi>10.1016/S0305-0491(00)00247-9</doi><tpages>10</tpages></addata></record> |
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subjects | Amino acid modifiers Animals Catalytic activity Chromatography, High Pressure Liquid Coproporphyria Coproporphyrin Coproporphyrinogen oxidase Coproporphyrinogen Oxidase - chemistry Coproporphyrinogen Oxidase - metabolism Coproporphyrinogen Oxidase - physiology Detergents - pharmacology Diethyl Pyrocarbonate - metabolism Enzyme Humans Liver - enzymology Octoxynol - pharmacology Phenylglyoxal - pharmacology Phospholipids - metabolism Polysorbates - pharmacology Protoporphyrin IX Protoporphyrinogen IX Purification Rat liver Rats Rats, Sprague-Dawley Water - metabolism |
title | Function and structure of rat hepatic coproporphyrinogen oxidase |
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