Different Recognition by Peroxisome Proliferator Structures in Rat Peroxisomal Induction : Application of Sandwich ELISA Using Monoclonal Antibody against Rat Peroxisomes
A novel assay for a peroxisomal β-oxidation enzyme by sandwich ELISA using a monoclonal antibody (RPX-5) against purified rat liver peroxisomes was developed. Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the β-oxidation pathw...
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| Veröffentlicht in: | Biological & pharmaceutical bulletin 2000/01/01, Vol.23(1), pp.12-16 |
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| creator | KAWANO, Hiroko NISHI, Fumihiro KAMITANI, Yuuko OCHI, Hitoshi MIYAKE, Masaharu MAYUMI, Tadanori HAMA, Takao |
| description | A novel assay for a peroxisomal β-oxidation enzyme by sandwich ELISA using a monoclonal antibody (RPX-5) against purified rat liver peroxisomes was developed. Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the β-oxidation pathway. Immunoprecipitation by RPX-5 and the resulting reduction of PBE activity were dependent on RPX-5 concentrations. Sandwich ELISA using RPX-5 could be used to assay PBE in the range of 30 to 2000 ng protein/ml. In rat hepatocyte cultures, the PBE amount by this assay correlated well with PBE activity, with correlation coefficients of 0.965. Studying the mechanisms of peroxisomal induction, patterns of peroxisomal induction were examined by co-treatment of rat hepatocytes with various peroxisome proliferators (PxPs). Treatment with clofibrate and bezafibrate resulted in neither an additive nor synergistic effect on PBE level. On the other hand, co-treatment with either bezafibrate-Wy-14, 643 or clofibrate-MEHP(mono(2-ethylhexyl)phthalate) both resulted in an additive effect. From these results, it is suggested that PxPs of the fibrate group may exert their functions via a common process, and non-fibrate PxPs via a different process in hepatocytes. The cognition site for peroxisome proliferators, therefore, might not involve a single site for inducing peroxisomal enzymes. |
| doi_str_mv | 10.1248/bpb.23.12 |
| format | Article |
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Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the β-oxidation pathway. Immunoprecipitation by RPX-5 and the resulting reduction of PBE activity were dependent on RPX-5 concentrations. Sandwich ELISA using RPX-5 could be used to assay PBE in the range of 30 to 2000 ng protein/ml. In rat hepatocyte cultures, the PBE amount by this assay correlated well with PBE activity, with correlation coefficients of 0.965. Studying the mechanisms of peroxisomal induction, patterns of peroxisomal induction were examined by co-treatment of rat hepatocytes with various peroxisome proliferators (PxPs). Treatment with clofibrate and bezafibrate resulted in neither an additive nor synergistic effect on PBE level. On the other hand, co-treatment with either bezafibrate-Wy-14, 643 or clofibrate-MEHP(mono(2-ethylhexyl)phthalate) both resulted in an additive effect. From these results, it is suggested that PxPs of the fibrate group may exert their functions via a common process, and non-fibrate PxPs via a different process in hepatocytes. The cognition site for peroxisome proliferators, therefore, might not involve a single site for inducing peroxisomal enzymes.</description><identifier>ISSN: 0918-6158</identifier><identifier>EISSN: 1347-5215</identifier><identifier>DOI: 10.1248/bpb.23.12</identifier><identifier>PMID: 10706403</identifier><language>eng</language><publisher>Tokyo: The Pharmaceutical Society of Japan</publisher><subject>3-Hydroxyacyl CoA Dehydrogenases - immunology ; 3-Hydroxyacyl CoA Dehydrogenases - metabolism ; Analysis ; Animals ; Antibodies, Monoclonal - biosynthesis ; Biological and medical sciences ; Cells, Cultured ; co-treatment ; Enoyl-CoA Hydratase - immunology ; Enoyl-CoA Hydratase - metabolism ; Enzyme-Linked Immunosorbent Assay - methods ; General pharmacology ; Isomerases ; Lipid Peroxidation ; Liver - enzymology ; Liver - metabolism ; Male ; Medical sciences ; Mice ; Mice, Inbred BALB C ; Multienzyme Complexes - immunology ; Multienzyme Complexes - metabolism ; Peroxisomal Bifunctional Enzyme ; peroxisomal β-oxidation ; peroxisome proliferator ; Peroxisome Proliferators - pharmacology ; Peroxisomes - enzymology ; Peroxisomes - immunology ; Peroxisomes - metabolism ; Pharmacology. Drug treatments ; rat hepatocyte ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear - metabolism ; sandwich ELISA ; Structure-Activity Relationship ; Transcription Factors - metabolism</subject><ispartof>Biological and Pharmaceutical Bulletin, 2000/01/01, Vol.23(1), pp.12-16</ispartof><rights>The Pharmaceutical Society of Japan</rights><rights>2000 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,1877,4010,27900,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1279481$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10706403$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KAWANO, Hiroko</creatorcontrib><creatorcontrib>NISHI, Fumihiro</creatorcontrib><creatorcontrib>KAMITANI, Yuuko</creatorcontrib><creatorcontrib>OCHI, Hitoshi</creatorcontrib><creatorcontrib>MIYAKE, Masaharu</creatorcontrib><creatorcontrib>MAYUMI, Tadanori</creatorcontrib><creatorcontrib>HAMA, Takao</creatorcontrib><title>Different Recognition by Peroxisome Proliferator Structures in Rat Peroxisomal Induction : Application of Sandwich ELISA Using Monoclonal Antibody against Rat Peroxisomes</title><title>Biological & pharmaceutical bulletin</title><addtitle>Biol Pharm Bull</addtitle><description>A novel assay for a peroxisomal β-oxidation enzyme by sandwich ELISA using a monoclonal antibody (RPX-5) against purified rat liver peroxisomes was developed. Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the β-oxidation pathway. Immunoprecipitation by RPX-5 and the resulting reduction of PBE activity were dependent on RPX-5 concentrations. Sandwich ELISA using RPX-5 could be used to assay PBE in the range of 30 to 2000 ng protein/ml. In rat hepatocyte cultures, the PBE amount by this assay correlated well with PBE activity, with correlation coefficients of 0.965. Studying the mechanisms of peroxisomal induction, patterns of peroxisomal induction were examined by co-treatment of rat hepatocytes with various peroxisome proliferators (PxPs). Treatment with clofibrate and bezafibrate resulted in neither an additive nor synergistic effect on PBE level. On the other hand, co-treatment with either bezafibrate-Wy-14, 643 or clofibrate-MEHP(mono(2-ethylhexyl)phthalate) both resulted in an additive effect. From these results, it is suggested that PxPs of the fibrate group may exert their functions via a common process, and non-fibrate PxPs via a different process in hepatocytes. The cognition site for peroxisome proliferators, therefore, might not involve a single site for inducing peroxisomal enzymes.</description><subject>3-Hydroxyacyl CoA Dehydrogenases - immunology</subject><subject>3-Hydroxyacyl CoA Dehydrogenases - metabolism</subject><subject>Analysis</subject><subject>Animals</subject><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>co-treatment</subject><subject>Enoyl-CoA Hydratase - immunology</subject><subject>Enoyl-CoA Hydratase - metabolism</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>General pharmacology</subject><subject>Isomerases</subject><subject>Lipid Peroxidation</subject><subject>Liver - enzymology</subject><subject>Liver - metabolism</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Multienzyme Complexes - immunology</subject><subject>Multienzyme Complexes - metabolism</subject><subject>Peroxisomal Bifunctional Enzyme</subject><subject>peroxisomal β-oxidation</subject><subject>peroxisome proliferator</subject><subject>Peroxisome Proliferators - pharmacology</subject><subject>Peroxisomes - enzymology</subject><subject>Peroxisomes - immunology</subject><subject>Peroxisomes - metabolism</subject><subject>Pharmacology. Drug treatments</subject><subject>rat hepatocyte</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Receptors, Cytoplasmic and Nuclear - metabolism</subject><subject>sandwich ELISA</subject><subject>Structure-Activity Relationship</subject><subject>Transcription Factors - metabolism</subject><issn>0918-6158</issn><issn>1347-5215</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkM1uEzEUhS0EoqGw4AWQF2xYTPHvZIZdWgpECqJqynp0x7lOXU3ske0I8ko8JU6namFzfazz3XOlQ8hbzs64UM3HfuzPhCz6GZlxqeaVFlw_JzPW8qaquW5OyKuU7hhjcybkS3LCi6gVkzPy57OzFiP6TK_RhK132QVP-wO9whh-uxR2SK9iGFyhIIdI1znuTd5HTNR5eg35iYSBLv2muMeIT3QxjoMzcP8Llq7Bb345c0svV8v1gv5Mzm_p9-CDGYIvqwufXR82BwpbcD7l_7MxvSYvLAwJ3zy8p-Tmy-XNxbdq9ePr8mKxqozWda76xqDRgELNFUcBtayxQWGsqnWrmGB1o8EqC8Yg8gbaXihtmVJG9YKBPCUfplgTQ0oRbTdGt4N46DjrjnV3pe5OyKIL-25ix32_w80_5NRvAd4_AJAMDDaCNy49cWLeqoYX7HzC7lKGLT76ELMzAx4v8raV91enwcWjaW4hdujlX6uUo2U</recordid><startdate>20000101</startdate><enddate>20000101</enddate><creator>KAWANO, Hiroko</creator><creator>NISHI, Fumihiro</creator><creator>KAMITANI, Yuuko</creator><creator>OCHI, Hitoshi</creator><creator>MIYAKE, Masaharu</creator><creator>MAYUMI, Tadanori</creator><creator>HAMA, Takao</creator><general>The Pharmaceutical Society of Japan</general><general>Maruzen</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20000101</creationdate><title>Different Recognition by Peroxisome Proliferator Structures in Rat Peroxisomal Induction : Application of Sandwich ELISA Using Monoclonal Antibody against Rat Peroxisomes</title><author>KAWANO, Hiroko ; NISHI, Fumihiro ; KAMITANI, Yuuko ; OCHI, Hitoshi ; MIYAKE, Masaharu ; MAYUMI, Tadanori ; HAMA, Takao</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c556t-b8cec5ae24741e2a636e8e2cf46594020685af4faccee18a9b245f044c4b20a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>3-Hydroxyacyl CoA Dehydrogenases - immunology</topic><topic>3-Hydroxyacyl CoA Dehydrogenases - metabolism</topic><topic>Analysis</topic><topic>Animals</topic><topic>Antibodies, Monoclonal - biosynthesis</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>co-treatment</topic><topic>Enoyl-CoA Hydratase - immunology</topic><topic>Enoyl-CoA Hydratase - metabolism</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>General pharmacology</topic><topic>Isomerases</topic><topic>Lipid Peroxidation</topic><topic>Liver - enzymology</topic><topic>Liver - metabolism</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Multienzyme Complexes - immunology</topic><topic>Multienzyme Complexes - metabolism</topic><topic>Peroxisomal Bifunctional Enzyme</topic><topic>peroxisomal β-oxidation</topic><topic>peroxisome proliferator</topic><topic>Peroxisome Proliferators - pharmacology</topic><topic>Peroxisomes - enzymology</topic><topic>Peroxisomes - immunology</topic><topic>Peroxisomes - metabolism</topic><topic>Pharmacology. Drug treatments</topic><topic>rat hepatocyte</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Receptors, Cytoplasmic and Nuclear - metabolism</topic><topic>sandwich ELISA</topic><topic>Structure-Activity Relationship</topic><topic>Transcription Factors - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KAWANO, Hiroko</creatorcontrib><creatorcontrib>NISHI, Fumihiro</creatorcontrib><creatorcontrib>KAMITANI, Yuuko</creatorcontrib><creatorcontrib>OCHI, Hitoshi</creatorcontrib><creatorcontrib>MIYAKE, Masaharu</creatorcontrib><creatorcontrib>MAYUMI, Tadanori</creatorcontrib><creatorcontrib>HAMA, Takao</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>Biological & pharmaceutical bulletin</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KAWANO, Hiroko</au><au>NISHI, Fumihiro</au><au>KAMITANI, Yuuko</au><au>OCHI, Hitoshi</au><au>MIYAKE, Masaharu</au><au>MAYUMI, Tadanori</au><au>HAMA, Takao</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Different Recognition by Peroxisome Proliferator Structures in Rat Peroxisomal Induction : Application of Sandwich ELISA Using Monoclonal Antibody against Rat Peroxisomes</atitle><jtitle>Biological & pharmaceutical bulletin</jtitle><addtitle>Biol Pharm Bull</addtitle><date>2000-01-01</date><risdate>2000</risdate><volume>23</volume><issue>1</issue><spage>12</spage><epage>16</epage><pages>12-16</pages><issn>0918-6158</issn><eissn>1347-5215</eissn><abstract>A novel assay for a peroxisomal β-oxidation enzyme by sandwich ELISA using a monoclonal antibody (RPX-5) against purified rat liver peroxisomes was developed. Immunoblot analysis revealed that RPX-5 recognized a 78 Kd protein, which is a peroxisomal bifunctional enzyme (PBE) in the β-oxidation pathway. Immunoprecipitation by RPX-5 and the resulting reduction of PBE activity were dependent on RPX-5 concentrations. Sandwich ELISA using RPX-5 could be used to assay PBE in the range of 30 to 2000 ng protein/ml. In rat hepatocyte cultures, the PBE amount by this assay correlated well with PBE activity, with correlation coefficients of 0.965. Studying the mechanisms of peroxisomal induction, patterns of peroxisomal induction were examined by co-treatment of rat hepatocytes with various peroxisome proliferators (PxPs). Treatment with clofibrate and bezafibrate resulted in neither an additive nor synergistic effect on PBE level. On the other hand, co-treatment with either bezafibrate-Wy-14, 643 or clofibrate-MEHP(mono(2-ethylhexyl)phthalate) both resulted in an additive effect. From these results, it is suggested that PxPs of the fibrate group may exert their functions via a common process, and non-fibrate PxPs via a different process in hepatocytes. The cognition site for peroxisome proliferators, therefore, might not involve a single site for inducing peroxisomal enzymes.</abstract><cop>Tokyo</cop><pub>The Pharmaceutical Society of Japan</pub><pmid>10706403</pmid><doi>10.1248/bpb.23.12</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| source | J-STAGE Free; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Free Full-Text Journals in Chemistry |
| subjects | 3-Hydroxyacyl CoA Dehydrogenases - immunology 3-Hydroxyacyl CoA Dehydrogenases - metabolism Analysis Animals Antibodies, Monoclonal - biosynthesis Biological and medical sciences Cells, Cultured co-treatment Enoyl-CoA Hydratase - immunology Enoyl-CoA Hydratase - metabolism Enzyme-Linked Immunosorbent Assay - methods General pharmacology Isomerases Lipid Peroxidation Liver - enzymology Liver - metabolism Male Medical sciences Mice Mice, Inbred BALB C Multienzyme Complexes - immunology Multienzyme Complexes - metabolism Peroxisomal Bifunctional Enzyme peroxisomal β-oxidation peroxisome proliferator Peroxisome Proliferators - pharmacology Peroxisomes - enzymology Peroxisomes - immunology Peroxisomes - metabolism Pharmacology. Drug treatments rat hepatocyte Rats Rats, Wistar Receptors, Cytoplasmic and Nuclear - metabolism sandwich ELISA Structure-Activity Relationship Transcription Factors - metabolism |
| title | Different Recognition by Peroxisome Proliferator Structures in Rat Peroxisomal Induction : Application of Sandwich ELISA Using Monoclonal Antibody against Rat Peroxisomes |
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