Subcellular Localization of Sterol Carrier Protein-2 in Rat Hepatocytes: Its Primary Localization to Peroxisomes
Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbi...
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| Veröffentlicht in: | The Journal of cell biology 1989-04, Vol.108 (4), p.1353-1361 |
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| container_title | The Journal of cell biology |
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| creator | Keller, G. A. Scallen, T. J. Clarke, D. Maher, P. A. Krisans, S. K. Singer, S. J. |
| description | Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbit polyclonal antibodies against electrophoretically pure SCP-2 from rat liver, we demonstrate by immunoelectron microscopic labeling of ultrathin frozen sections of rat liver that the largest concentration of SCP-2 is inside peroxisomes. In addition the immunolabeling indicates that there are significant concentrations of SCP-2 inside mitochondria, and associated with the endoplasmic reticulum and the cytosol, but not inside the Golgi apparatus, lysosomes, or the nucleus. These results were confirmed by immunoblotting experiments with proteins from purified subcellular fractions of the rat liver cells carried out with the anti-SCP-2 antibodies. The large concentration of SCP-2 inside peroxisomes strongly supports the proposal that peroxisomes are critical sites of cholesterol utilization and biosynthesis. The presence of SCP-2 inside peroxisomes and mitochondria raises questions about the mechanisms involved in the differential targeting of SCP-2 to these organelles. |
| doi_str_mv | 10.1083/jcb.108.4.1353 |
| format | Article |
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A. ; Scallen, T. J. ; Clarke, D. ; Maher, P. A. ; Krisans, S. K. ; Singer, S. J.</creator><creatorcontrib>Keller, G. A. ; Scallen, T. J. ; Clarke, D. ; Maher, P. A. ; Krisans, S. K. ; Singer, S. J.</creatorcontrib><description>Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbit polyclonal antibodies against electrophoretically pure SCP-2 from rat liver, we demonstrate by immunoelectron microscopic labeling of ultrathin frozen sections of rat liver that the largest concentration of SCP-2 is inside peroxisomes. In addition the immunolabeling indicates that there are significant concentrations of SCP-2 inside mitochondria, and associated with the endoplasmic reticulum and the cytosol, but not inside the Golgi apparatus, lysosomes, or the nucleus. These results were confirmed by immunoblotting experiments with proteins from purified subcellular fractions of the rat liver cells carried out with the anti-SCP-2 antibodies. The large concentration of SCP-2 inside peroxisomes strongly supports the proposal that peroxisomes are critical sites of cholesterol utilization and biosynthesis. The presence of SCP-2 inside peroxisomes and mitochondria raises questions about the mechanisms involved in the differential targeting of SCP-2 to these organelles.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.108.4.1353</identifier><identifier>PMID: 2925789</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>New York, NY: Rockefeller University Press</publisher><subject>Analytical, structural and metabolic biochemistry ; Animals ; Antibodies ; Binding and carrier proteins ; Biological and medical sciences ; Carrier proteins ; Carrier Proteins - immunology ; Carrier Proteins - isolation & purification ; Carrier Proteins - metabolism ; Cholesterols ; Cytosol ; Electrophoresis, Polyacrylamide Gel ; Frozen sections ; Fundamental and applied biological sciences. Psychology ; hepatocytes ; Immunoblotting ; Immunohistochemistry ; Liver ; Liver - metabolism ; Liver - ultrastructure ; Microbodies - metabolism ; Microbodies - ultrastructure ; Microscopy, Electron ; Microsomes, Liver - metabolism ; Microsomes, Liver - ultrastructure ; Mitochondria ; Mitochondria, Liver - metabolism ; Mitochondria, Liver - ultrastructure ; Molecular Weight ; Peroxisomes ; Plant Proteins ; Proteins ; Rats ; Rats, Inbred Strains ; Sterols ; Sterols - metabolism</subject><ispartof>The Journal of cell biology, 1989-04, Vol.108 (4), p.1353-1361</ispartof><rights>Copyright 1989 The Rockefeller University Press</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c532t-1335e4ca45394015184fe0a6b46d31ff797f7c8fdda9bc97e20db8c8d591e2e93</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115497/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115497/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,886,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7256562$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2925789$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Keller, G. A.</creatorcontrib><creatorcontrib>Scallen, T. J.</creatorcontrib><creatorcontrib>Clarke, D.</creatorcontrib><creatorcontrib>Maher, P. A.</creatorcontrib><creatorcontrib>Krisans, S. K.</creatorcontrib><creatorcontrib>Singer, S. J.</creatorcontrib><title>Subcellular Localization of Sterol Carrier Protein-2 in Rat Hepatocytes: Its Primary Localization to Peroxisomes</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbit polyclonal antibodies against electrophoretically pure SCP-2 from rat liver, we demonstrate by immunoelectron microscopic labeling of ultrathin frozen sections of rat liver that the largest concentration of SCP-2 is inside peroxisomes. In addition the immunolabeling indicates that there are significant concentrations of SCP-2 inside mitochondria, and associated with the endoplasmic reticulum and the cytosol, but not inside the Golgi apparatus, lysosomes, or the nucleus. These results were confirmed by immunoblotting experiments with proteins from purified subcellular fractions of the rat liver cells carried out with the anti-SCP-2 antibodies. The large concentration of SCP-2 inside peroxisomes strongly supports the proposal that peroxisomes are critical sites of cholesterol utilization and biosynthesis. The presence of SCP-2 inside peroxisomes and mitochondria raises questions about the mechanisms involved in the differential targeting of SCP-2 to these organelles.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Binding and carrier proteins</subject><subject>Biological and medical sciences</subject><subject>Carrier proteins</subject><subject>Carrier Proteins - immunology</subject><subject>Carrier Proteins - isolation & purification</subject><subject>Carrier Proteins - metabolism</subject><subject>Cholesterols</subject><subject>Cytosol</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Frozen sections</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>hepatocytes</subject><subject>Immunoblotting</subject><subject>Immunohistochemistry</subject><subject>Liver</subject><subject>Liver - metabolism</subject><subject>Liver - ultrastructure</subject><subject>Microbodies - metabolism</subject><subject>Microbodies - ultrastructure</subject><subject>Microscopy, Electron</subject><subject>Microsomes, Liver - metabolism</subject><subject>Microsomes, Liver - ultrastructure</subject><subject>Mitochondria</subject><subject>Mitochondria, Liver - metabolism</subject><subject>Mitochondria, Liver - ultrastructure</subject><subject>Molecular Weight</subject><subject>Peroxisomes</subject><subject>Plant Proteins</subject><subject>Proteins</subject><subject>Rats</subject><subject>Rats, Inbred Strains</subject><subject>Sterols</subject><subject>Sterols - metabolism</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9rFDEcxYModa1ePSnkIL3Nmp-TxENBlmoLCy1WzyGTSTTL7GRNMqX1rzfDLqs99ZQvvE9evnkPgLcYLTGS9OPGdvOwZEtMOX0GFpgz1EjM0HOwQIjgRnHCX4JXOW8QQkwwegJOiCJcSLUAu9ups24YpsEkuI7WDOGPKSGOMHp4W1yKA1yZlIJL8CbF4sLYEBhG-M0UeOl2pkT7UFz-BK9KrkTYmvTw2KhEeFN97kOOW5dfgxfeDNm9OZyn4MeXi--ry2Z9_fVq9XndWE5JaTCl3DFrGKeKIcyxZN4h03as7Sn2XijhhZW-743qrBKOoL6TVvZcYUecoqfgfO-7m7qt660bSzKD3u031NEE_VgZwy_9M95pgmuCSlSDs4NBir8nl4vehjxHZUYXp6yFlJK2mD0JYk5Qqwiq4HIP2hRzTs4ft8FIz2XqWuY8aKbnMuuF9___4Ygf2qv6h4Nucg3cJzPakI-YILzlLanYuz22ySWmf4-2NWRG6V9Y6bNU</recordid><startdate>19890401</startdate><enddate>19890401</enddate><creator>Keller, G. A.</creator><creator>Scallen, T. J.</creator><creator>Clarke, D.</creator><creator>Maher, P. A.</creator><creator>Krisans, S. K.</creator><creator>Singer, S. J.</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</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><scope>8FD</scope><scope>FR3</scope><scope>M7Z</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19890401</creationdate><title>Subcellular Localization of Sterol Carrier Protein-2 in Rat Hepatocytes: Its Primary Localization to Peroxisomes</title><author>Keller, G. A. ; Scallen, T. J. ; Clarke, D. ; Maher, P. A. ; Krisans, S. K. ; Singer, S. J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c532t-1335e4ca45394015184fe0a6b46d31ff797f7c8fdda9bc97e20db8c8d591e2e93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Binding and carrier proteins</topic><topic>Biological and medical sciences</topic><topic>Carrier proteins</topic><topic>Carrier Proteins - immunology</topic><topic>Carrier Proteins - isolation & purification</topic><topic>Carrier Proteins - metabolism</topic><topic>Cholesterols</topic><topic>Cytosol</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Frozen sections</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>hepatocytes</topic><topic>Immunoblotting</topic><topic>Immunohistochemistry</topic><topic>Liver</topic><topic>Liver - metabolism</topic><topic>Liver - ultrastructure</topic><topic>Microbodies - metabolism</topic><topic>Microbodies - ultrastructure</topic><topic>Microscopy, Electron</topic><topic>Microsomes, Liver - metabolism</topic><topic>Microsomes, Liver - ultrastructure</topic><topic>Mitochondria</topic><topic>Mitochondria, Liver - metabolism</topic><topic>Mitochondria, Liver - ultrastructure</topic><topic>Molecular Weight</topic><topic>Peroxisomes</topic><topic>Plant Proteins</topic><topic>Proteins</topic><topic>Rats</topic><topic>Rats, Inbred Strains</topic><topic>Sterols</topic><topic>Sterols - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Keller, G. A.</creatorcontrib><creatorcontrib>Scallen, T. J.</creatorcontrib><creatorcontrib>Clarke, D.</creatorcontrib><creatorcontrib>Maher, P. A.</creatorcontrib><creatorcontrib>Krisans, S. K.</creatorcontrib><creatorcontrib>Singer, S. J.</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><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Keller, G. A.</au><au>Scallen, T. J.</au><au>Clarke, D.</au><au>Maher, P. A.</au><au>Krisans, S. K.</au><au>Singer, S. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Subcellular Localization of Sterol Carrier Protein-2 in Rat Hepatocytes: Its Primary Localization to Peroxisomes</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1989-04-01</date><risdate>1989</risdate><volume>108</volume><issue>4</issue><spage>1353</spage><epage>1361</epage><pages>1353-1361</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>Sterol carrier protein-2 (SCP-2) is a nonenzymatic protein of 13.5 kD which has been shown in in vitro experiments to be required for several stages in cholesterol utilization and biosynthesis. The subcellular localization of SCP-2 has not been definitively established. Using affinity-purified rabbit polyclonal antibodies against electrophoretically pure SCP-2 from rat liver, we demonstrate by immunoelectron microscopic labeling of ultrathin frozen sections of rat liver that the largest concentration of SCP-2 is inside peroxisomes. In addition the immunolabeling indicates that there are significant concentrations of SCP-2 inside mitochondria, and associated with the endoplasmic reticulum and the cytosol, but not inside the Golgi apparatus, lysosomes, or the nucleus. These results were confirmed by immunoblotting experiments with proteins from purified subcellular fractions of the rat liver cells carried out with the anti-SCP-2 antibodies. The large concentration of SCP-2 inside peroxisomes strongly supports the proposal that peroxisomes are critical sites of cholesterol utilization and biosynthesis. The presence of SCP-2 inside peroxisomes and mitochondria raises questions about the mechanisms involved in the differential targeting of SCP-2 to these organelles.</abstract><cop>New York, NY</cop><pub>Rockefeller University Press</pub><pmid>2925789</pmid><doi>10.1083/jcb.108.4.1353</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of cell biology, 1989-04, Vol.108 (4), p.1353-1361 |
| issn | 0021-9525 1540-8140 |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Analytical, structural and metabolic biochemistry Animals Antibodies Binding and carrier proteins Biological and medical sciences Carrier proteins Carrier Proteins - immunology Carrier Proteins - isolation & purification Carrier Proteins - metabolism Cholesterols Cytosol Electrophoresis, Polyacrylamide Gel Frozen sections Fundamental and applied biological sciences. Psychology hepatocytes Immunoblotting Immunohistochemistry Liver Liver - metabolism Liver - ultrastructure Microbodies - metabolism Microbodies - ultrastructure Microscopy, Electron Microsomes, Liver - metabolism Microsomes, Liver - ultrastructure Mitochondria Mitochondria, Liver - metabolism Mitochondria, Liver - ultrastructure Molecular Weight Peroxisomes Plant Proteins Proteins Rats Rats, Inbred Strains Sterols Sterols - metabolism |
| title | Subcellular Localization of Sterol Carrier Protein-2 in Rat Hepatocytes: Its Primary Localization to Peroxisomes |
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