Cultured Human Fibroblasts Synthesize and Secrete Thrombospondin and Incorporate It into Extracellular Matrix
Thrombospondin, a major glycoprotein released from α granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture mediu...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1983-02, Vol.80 (4), p.998-1002 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Jaffe, Eric A. Ruggiero, Joseph T. Lawrence L. K. Leung Doyle, Mary Jean McKeown-Longo, Paula J. Mosher, Deane F. |
| description | Thrombospondin, a major glycoprotein released from α granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 μ g per 106 cells per 24 hr, respectively); secretion was blocked by cycloheximide. [3H]Thrombospondin was isolated from [3H]leucine-labeled fibroblast postculture medium and from cell layers with rabbit polyclonal or mouse monoclonal anti-thrombospondin coupled to staphylococcal protein A-Sepharose. The immunologically isolated [3H]thrombospondin migrated in NaDodSO4/polyacrylamide gels with purified marker platelet thrombospondin both with and without reduction. Immunofluorescence microscopy using rabbit polyclonal and mouse monoclonal anti-thrombospondin antibodies localized thrombospondin to the fibrillar extracellular matrix surrounding the cells. Thus, cultured human fibroblasts secrete thrombospondin and incorporate it into the extracellular matrix. |
| doi_str_mv | 10.1073/pnas.80.4.998 |
| format | Article |
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K. Leung ; Doyle, Mary Jean ; McKeown-Longo, Paula J. ; Mosher, Deane F.</creator><creatorcontrib>Jaffe, Eric A. ; Ruggiero, Joseph T. ; Lawrence L. K. Leung ; Doyle, Mary Jean ; McKeown-Longo, Paula J. ; Mosher, Deane F.</creatorcontrib><description>Thrombospondin, a major glycoprotein released from α granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 μ g per 106 cells per 24 hr, respectively); secretion was blocked by cycloheximide. [3H]Thrombospondin was isolated from [3H]leucine-labeled fibroblast postculture medium and from cell layers with rabbit polyclonal or mouse monoclonal anti-thrombospondin coupled to staphylococcal protein A-Sepharose. The immunologically isolated [3H]thrombospondin migrated in NaDodSO4/polyacrylamide gels with purified marker platelet thrombospondin both with and without reduction. Immunofluorescence microscopy using rabbit polyclonal and mouse monoclonal anti-thrombospondin antibodies localized thrombospondin to the fibrillar extracellular matrix surrounding the cells. Thus, cultured human fibroblasts secrete thrombospondin and incorporate it into the extracellular matrix.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.80.4.998</identifier><identifier>PMID: 6341993</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Antibodies ; Cells, Cultured ; Cultured cells ; Endothelial cells ; Extracellular matrix ; Extracellular Space - physiology ; Fibroblasts ; Fibroblasts - metabolism ; Fluorescent Antibody Technique ; Foreskin ; Glycoproteins - metabolism ; Humans ; Kinetics ; Lungs ; Male ; Platelets ; Rabbits ; Thrombospondins</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1983-02, Vol.80 (4), p.998-1002</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c451t-541e17ac75d017c86e5344395180f77ac5ffc5203bbc5164a40abd9aa9c42bce3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/80/4.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/13870$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/13870$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27924,27925,53791,53793,58017,58250</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6341993$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jaffe, Eric A.</creatorcontrib><creatorcontrib>Ruggiero, Joseph T.</creatorcontrib><creatorcontrib>Lawrence L. K. Leung</creatorcontrib><creatorcontrib>Doyle, Mary Jean</creatorcontrib><creatorcontrib>McKeown-Longo, Paula J.</creatorcontrib><creatorcontrib>Mosher, Deane F.</creatorcontrib><title>Cultured Human Fibroblasts Synthesize and Secrete Thrombospondin and Incorporate It into Extracellular Matrix</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Thrombospondin, a major glycoprotein released from α granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 μ g per 106 cells per 24 hr, respectively); secretion was blocked by cycloheximide. [3H]Thrombospondin was isolated from [3H]leucine-labeled fibroblast postculture medium and from cell layers with rabbit polyclonal or mouse monoclonal anti-thrombospondin coupled to staphylococcal protein A-Sepharose. The immunologically isolated [3H]thrombospondin migrated in NaDodSO4/polyacrylamide gels with purified marker platelet thrombospondin both with and without reduction. Immunofluorescence microscopy using rabbit polyclonal and mouse monoclonal anti-thrombospondin antibodies localized thrombospondin to the fibrillar extracellular matrix surrounding the cells. Thus, cultured human fibroblasts secrete thrombospondin and incorporate it into the extracellular matrix.</description><subject>Antibodies</subject><subject>Cells, Cultured</subject><subject>Cultured cells</subject><subject>Endothelial cells</subject><subject>Extracellular matrix</subject><subject>Extracellular Space - physiology</subject><subject>Fibroblasts</subject><subject>Fibroblasts - metabolism</subject><subject>Fluorescent Antibody Technique</subject><subject>Foreskin</subject><subject>Glycoproteins - metabolism</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Lungs</subject><subject>Male</subject><subject>Platelets</subject><subject>Rabbits</subject><subject>Thrombospondins</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1983</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc9vFCEYhonR1G316MVowkVvs34sMDMcPJhNf2xS46H1TBiGcWkYGIExW_96WXet9eKJw_O88JIXoVcElgQa-mHyKi1bWLKlEO0TtCAgSFUzAU_RAmDVVC1bsefoNKU7ABC8hRN0UlNGhKALNK5nl-doenw1j8rjC9vF0DmVcsI39z5vTbI_DVa-xzdGR5MNvt3GMHYhTcH31v9GG69DnEJUBW8ytj4HfL7LUWnj3OxUxJ9Vjnb3Aj0blEvm5fE8Q18vzm_XV9X1l8vN-tN1pRknueKMGNIo3fAeSKPb2nDKGBWctDA0BfBh0HwFtOs0JzVTDFTXC6WEZqtOG3qGPh7uneZuNL02vnRxcop2VPFeBmXlv8TbrfwWfkgqKCe85N8f8zF8n03KcrRp_xflTZiTbIHVFAgUsTqIOoaUohke3iAg9_PI_TzFl0yWeYr_9nGxB_u4R-Fvjnwf-0Mfxd_9B8thdi6bXS7e64N3l3KIfzvRtgH6C1pWr2A</recordid><startdate>19830201</startdate><enddate>19830201</enddate><creator>Jaffe, Eric A.</creator><creator>Ruggiero, Joseph T.</creator><creator>Lawrence L. K. Leung</creator><creator>Doyle, Mary Jean</creator><creator>McKeown-Longo, Paula J.</creator><creator>Mosher, Deane F.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</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><scope>5PM</scope></search><sort><creationdate>19830201</creationdate><title>Cultured Human Fibroblasts Synthesize and Secrete Thrombospondin and Incorporate It into Extracellular Matrix</title><author>Jaffe, Eric A. ; Ruggiero, Joseph T. ; Lawrence L. K. Leung ; Doyle, Mary Jean ; McKeown-Longo, Paula J. ; Mosher, Deane F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c451t-541e17ac75d017c86e5344395180f77ac5ffc5203bbc5164a40abd9aa9c42bce3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1983</creationdate><topic>Antibodies</topic><topic>Cells, Cultured</topic><topic>Cultured cells</topic><topic>Endothelial cells</topic><topic>Extracellular matrix</topic><topic>Extracellular Space - physiology</topic><topic>Fibroblasts</topic><topic>Fibroblasts - metabolism</topic><topic>Fluorescent Antibody Technique</topic><topic>Foreskin</topic><topic>Glycoproteins - metabolism</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Lungs</topic><topic>Male</topic><topic>Platelets</topic><topic>Rabbits</topic><topic>Thrombospondins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jaffe, Eric A.</creatorcontrib><creatorcontrib>Ruggiero, Joseph T.</creatorcontrib><creatorcontrib>Lawrence L. K. Leung</creatorcontrib><creatorcontrib>Doyle, Mary Jean</creatorcontrib><creatorcontrib>McKeown-Longo, Paula J.</creatorcontrib><creatorcontrib>Mosher, Deane F.</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><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jaffe, Eric A.</au><au>Ruggiero, Joseph T.</au><au>Lawrence L. K. Leung</au><au>Doyle, Mary Jean</au><au>McKeown-Longo, Paula J.</au><au>Mosher, Deane F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cultured Human Fibroblasts Synthesize and Secrete Thrombospondin and Incorporate It into Extracellular Matrix</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1983-02-01</date><risdate>1983</risdate><volume>80</volume><issue>4</issue><spage>998</spage><epage>1002</epage><pages>998-1002</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Thrombospondin, a major glycoprotein released from α granules of thrombin-stimulated platelets, is a disulfide-bonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 μ g per 106 cells per 24 hr, respectively); secretion was blocked by cycloheximide. [3H]Thrombospondin was isolated from [3H]leucine-labeled fibroblast postculture medium and from cell layers with rabbit polyclonal or mouse monoclonal anti-thrombospondin coupled to staphylococcal protein A-Sepharose. The immunologically isolated [3H]thrombospondin migrated in NaDodSO4/polyacrylamide gels with purified marker platelet thrombospondin both with and without reduction. Immunofluorescence microscopy using rabbit polyclonal and mouse monoclonal anti-thrombospondin antibodies localized thrombospondin to the fibrillar extracellular matrix surrounding the cells. Thus, cultured human fibroblasts secrete thrombospondin and incorporate it into the extracellular matrix.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>6341993</pmid><doi>10.1073/pnas.80.4.998</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1983-02, Vol.80 (4), p.998-1002 |
| issn | 0027-8424 1091-6490 |
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| source | MEDLINE; JSTOR Archive Collection A-Z Listing; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Antibodies Cells, Cultured Cultured cells Endothelial cells Extracellular matrix Extracellular Space - physiology Fibroblasts Fibroblasts - metabolism Fluorescent Antibody Technique Foreskin Glycoproteins - metabolism Humans Kinetics Lungs Male Platelets Rabbits Thrombospondins |
| title | Cultured Human Fibroblasts Synthesize and Secrete Thrombospondin and Incorporate It into Extracellular Matrix |
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