Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues
A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecula...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 2003-12, Vol.278 (50), p.50546-50553 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 50553 |
|---|---|
| container_issue | 50 |
| container_start_page | 50546 |
| container_title | The Journal of biological chemistry |
| container_volume | 278 |
| creator | Kozaki, Tomohiro Matsui, Yoshito Gu, Jianguo Nishiuchi, Ryoko Sugiura, Nobuo Kimata, Koji Ozono, Keiichi Yoshikawa, Hideki Sekiguchi, Kiyotoshi |
| description | A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecular and biological properties. Although FNs have been shown to be secreted as disulfide-bonded dimers, cartilage-type FN was secreted mainly as a monomer. It was less potent than plasma-type FN in promoting cell adhesion and binding to integrin α5β1, although it was more active than plasma-type FN in binding to chondroitin sulfate E. When added exogenously, cartilage-type FN was poorly assembled into the fibrillar FN matrix, mostly because of its monomeric structure. Given that cartilage is characterized by its non-fibrillar matrix with abundant chondroitin sulfate-containing proteoglycans, it is likely that cartilage-type FN has evolved to adapt itself to the non-fibrillar structure of the cartilage matrix through acquisition of a novel mechanism of alternative pre-mRNA splicing. |
| doi_str_mv | 10.1074/jbc.M307432200 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_19202767</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925820755766</els_id><sourcerecordid>19202767</sourcerecordid><originalsourceid>FETCH-LOGICAL-c550t-85f9d7ba2f27e7940491f603826c775396e9882d12a7c2a91af20f82d8daa41b3</originalsourceid><addsrcrecordid>eNp1kMFrFDEUxoModq1ePUoO4m3WJDOZJEdZWi1UBangLbzJvHRTZpI1mW3Vv94su9KTIZDw8fve-_gIec3ZmjPVvb8b3PpzW3-tEIw9ISvOdNu0kv94SlaMCd4YIfUZeVHKHaunM_w5OeOdFNIYtSLzN3RpHkKEuNCLX7uMpYQUKcSRbraQwS2Ywx9YDmLyFOiXdI8TvQxDThHdEiK9KsmnPP-z40iruIG8hAluQ0z7Qm9CKXssL8kzD1PBV6f3nHy_vLjZfGquv3682ny4bpyUbGm09GZUAwgvFCrTHVL7nrVa9E4p2ZoejdZi5AKUE2A4eMF8FfQI0PGhPSfvjnN3Of2sexc7h-JwmiBijWO5EUyoXlVwfQRdTqVk9HaXwwz5t-XMHgq2tWD7WHA1vDlN3g8zjo_4qdEKvD0C23C7fQgZ7RCS2-JshdJWsnpl11dMHzGsNdwHzLa4gNHhWC1usWMK_4vwF1_Dlm4</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>19202767</pqid></control><display><type>article</type><title>Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Kozaki, Tomohiro ; Matsui, Yoshito ; Gu, Jianguo ; Nishiuchi, Ryoko ; Sugiura, Nobuo ; Kimata, Koji ; Ozono, Keiichi ; Yoshikawa, Hideki ; Sekiguchi, Kiyotoshi</creator><creatorcontrib>Kozaki, Tomohiro ; Matsui, Yoshito ; Gu, Jianguo ; Nishiuchi, Ryoko ; Sugiura, Nobuo ; Kimata, Koji ; Ozono, Keiichi ; Yoshikawa, Hideki ; Sekiguchi, Kiyotoshi</creatorcontrib><description>A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecular and biological properties. Although FNs have been shown to be secreted as disulfide-bonded dimers, cartilage-type FN was secreted mainly as a monomer. It was less potent than plasma-type FN in promoting cell adhesion and binding to integrin α5β1, although it was more active than plasma-type FN in binding to chondroitin sulfate E. When added exogenously, cartilage-type FN was poorly assembled into the fibrillar FN matrix, mostly because of its monomeric structure. Given that cartilage is characterized by its non-fibrillar matrix with abundant chondroitin sulfate-containing proteoglycans, it is likely that cartilage-type FN has evolved to adapt itself to the non-fibrillar structure of the cartilage matrix through acquisition of a novel mechanism of alternative pre-mRNA splicing.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M307432200</identifier><identifier>PMID: 14525997</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Antibodies, Monoclonal - metabolism ; Cartilage - metabolism ; Cell Adhesion ; Cell Line ; Cells, Cultured ; CHO Cells ; Chondroitin Sulfates - chemistry ; Cricetinae ; Disulfides - chemistry ; DNA, Complementary - metabolism ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Fibronectins - biosynthesis ; Fibronectins - chemistry ; Fluorescent Antibody Technique, Indirect ; Glycosaminoglycans - metabolism ; Heparin - metabolism ; Immunoblotting ; Integrin alpha5beta1 - metabolism ; Mice ; Models, Genetic ; Peptides - chemistry ; Protein Binding ; Protein Isoforms ; Proteoglycans - metabolism ; Recombinant Proteins - chemistry ; RNA - metabolism ; RNA Splicing ; Transfection</subject><ispartof>The Journal of biological chemistry, 2003-12, Vol.278 (50), p.50546-50553</ispartof><rights>2003 © 2003 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c550t-85f9d7ba2f27e7940491f603826c775396e9882d12a7c2a91af20f82d8daa41b3</citedby><cites>FETCH-LOGICAL-c550t-85f9d7ba2f27e7940491f603826c775396e9882d12a7c2a91af20f82d8daa41b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14525997$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kozaki, Tomohiro</creatorcontrib><creatorcontrib>Matsui, Yoshito</creatorcontrib><creatorcontrib>Gu, Jianguo</creatorcontrib><creatorcontrib>Nishiuchi, Ryoko</creatorcontrib><creatorcontrib>Sugiura, Nobuo</creatorcontrib><creatorcontrib>Kimata, Koji</creatorcontrib><creatorcontrib>Ozono, Keiichi</creatorcontrib><creatorcontrib>Yoshikawa, Hideki</creatorcontrib><creatorcontrib>Sekiguchi, Kiyotoshi</creatorcontrib><title>Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecular and biological properties. Although FNs have been shown to be secreted as disulfide-bonded dimers, cartilage-type FN was secreted mainly as a monomer. It was less potent than plasma-type FN in promoting cell adhesion and binding to integrin α5β1, although it was more active than plasma-type FN in binding to chondroitin sulfate E. When added exogenously, cartilage-type FN was poorly assembled into the fibrillar FN matrix, mostly because of its monomeric structure. Given that cartilage is characterized by its non-fibrillar matrix with abundant chondroitin sulfate-containing proteoglycans, it is likely that cartilage-type FN has evolved to adapt itself to the non-fibrillar structure of the cartilage matrix through acquisition of a novel mechanism of alternative pre-mRNA splicing.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - metabolism</subject><subject>Cartilage - metabolism</subject><subject>Cell Adhesion</subject><subject>Cell Line</subject><subject>Cells, Cultured</subject><subject>CHO Cells</subject><subject>Chondroitin Sulfates - chemistry</subject><subject>Cricetinae</subject><subject>Disulfides - chemistry</subject><subject>DNA, Complementary - metabolism</subject><subject>Dose-Response Relationship, Drug</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Fibronectins - biosynthesis</subject><subject>Fibronectins - chemistry</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Glycosaminoglycans - metabolism</subject><subject>Heparin - metabolism</subject><subject>Immunoblotting</subject><subject>Integrin alpha5beta1 - metabolism</subject><subject>Mice</subject><subject>Models, Genetic</subject><subject>Peptides - chemistry</subject><subject>Protein Binding</subject><subject>Protein Isoforms</subject><subject>Proteoglycans - metabolism</subject><subject>Recombinant Proteins - chemistry</subject><subject>RNA - metabolism</subject><subject>RNA Splicing</subject><subject>Transfection</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kMFrFDEUxoModq1ePUoO4m3WJDOZJEdZWi1UBangLbzJvHRTZpI1mW3Vv94su9KTIZDw8fve-_gIec3ZmjPVvb8b3PpzW3-tEIw9ISvOdNu0kv94SlaMCd4YIfUZeVHKHaunM_w5OeOdFNIYtSLzN3RpHkKEuNCLX7uMpYQUKcSRbraQwS2Ywx9YDmLyFOiXdI8TvQxDThHdEiK9KsmnPP-z40iruIG8hAluQ0z7Qm9CKXssL8kzD1PBV6f3nHy_vLjZfGquv3682ny4bpyUbGm09GZUAwgvFCrTHVL7nrVa9E4p2ZoejdZi5AKUE2A4eMF8FfQI0PGhPSfvjnN3Of2sexc7h-JwmiBijWO5EUyoXlVwfQRdTqVk9HaXwwz5t-XMHgq2tWD7WHA1vDlN3g8zjo_4qdEKvD0C23C7fQgZ7RCS2-JshdJWsnpl11dMHzGsNdwHzLa4gNHhWC1usWMK_4vwF1_Dlm4</recordid><startdate>20031212</startdate><enddate>20031212</enddate><creator>Kozaki, Tomohiro</creator><creator>Matsui, Yoshito</creator><creator>Gu, Jianguo</creator><creator>Nishiuchi, Ryoko</creator><creator>Sugiura, Nobuo</creator><creator>Kimata, Koji</creator><creator>Ozono, Keiichi</creator><creator>Yoshikawa, Hideki</creator><creator>Sekiguchi, Kiyotoshi</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</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>7QP</scope><scope>7TM</scope></search><sort><creationdate>20031212</creationdate><title>Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues</title><author>Kozaki, Tomohiro ; Matsui, Yoshito ; Gu, Jianguo ; Nishiuchi, Ryoko ; Sugiura, Nobuo ; Kimata, Koji ; Ozono, Keiichi ; Yoshikawa, Hideki ; Sekiguchi, Kiyotoshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c550t-85f9d7ba2f27e7940491f603826c775396e9882d12a7c2a91af20f82d8daa41b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - metabolism</topic><topic>Cartilage - metabolism</topic><topic>Cell Adhesion</topic><topic>Cell Line</topic><topic>Cells, Cultured</topic><topic>CHO Cells</topic><topic>Chondroitin Sulfates - chemistry</topic><topic>Cricetinae</topic><topic>Disulfides - chemistry</topic><topic>DNA, Complementary - metabolism</topic><topic>Dose-Response Relationship, Drug</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Fibronectins - biosynthesis</topic><topic>Fibronectins - chemistry</topic><topic>Fluorescent Antibody Technique, Indirect</topic><topic>Glycosaminoglycans - metabolism</topic><topic>Heparin - metabolism</topic><topic>Immunoblotting</topic><topic>Integrin alpha5beta1 - metabolism</topic><topic>Mice</topic><topic>Models, Genetic</topic><topic>Peptides - chemistry</topic><topic>Protein Binding</topic><topic>Protein Isoforms</topic><topic>Proteoglycans - metabolism</topic><topic>Recombinant Proteins - chemistry</topic><topic>RNA - metabolism</topic><topic>RNA Splicing</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kozaki, Tomohiro</creatorcontrib><creatorcontrib>Matsui, Yoshito</creatorcontrib><creatorcontrib>Gu, Jianguo</creatorcontrib><creatorcontrib>Nishiuchi, Ryoko</creatorcontrib><creatorcontrib>Sugiura, Nobuo</creatorcontrib><creatorcontrib>Kimata, Koji</creatorcontrib><creatorcontrib>Ozono, Keiichi</creatorcontrib><creatorcontrib>Yoshikawa, Hideki</creatorcontrib><creatorcontrib>Sekiguchi, Kiyotoshi</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Nucleic Acids Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kozaki, Tomohiro</au><au>Matsui, Yoshito</au><au>Gu, Jianguo</au><au>Nishiuchi, Ryoko</au><au>Sugiura, Nobuo</au><au>Kimata, Koji</au><au>Ozono, Keiichi</au><au>Yoshikawa, Hideki</au><au>Sekiguchi, Kiyotoshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2003-12-12</date><risdate>2003</risdate><volume>278</volume><issue>50</issue><spage>50546</spage><epage>50553</epage><pages>50546-50553</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>A novel fibronectin (FN) isoform lacking the segment from IIICS (type III connecting segment) through the I-10 module is expressed predominantly in normal cartilaginous tissues. We expressed and purified recombinant cartilage-type FN using a mammalian expression system and characterized its molecular and biological properties. Although FNs have been shown to be secreted as disulfide-bonded dimers, cartilage-type FN was secreted mainly as a monomer. It was less potent than plasma-type FN in promoting cell adhesion and binding to integrin α5β1, although it was more active than plasma-type FN in binding to chondroitin sulfate E. When added exogenously, cartilage-type FN was poorly assembled into the fibrillar FN matrix, mostly because of its monomeric structure. Given that cartilage is characterized by its non-fibrillar matrix with abundant chondroitin sulfate-containing proteoglycans, it is likely that cartilage-type FN has evolved to adapt itself to the non-fibrillar structure of the cartilage matrix through acquisition of a novel mechanism of alternative pre-mRNA splicing.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>14525997</pmid><doi>10.1074/jbc.M307432200</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 2003-12, Vol.278 (50), p.50546-50553 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_19202767 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animals Antibodies, Monoclonal - metabolism Cartilage - metabolism Cell Adhesion Cell Line Cells, Cultured CHO Cells Chondroitin Sulfates - chemistry Cricetinae Disulfides - chemistry DNA, Complementary - metabolism Dose-Response Relationship, Drug Electrophoresis, Polyacrylamide Gel Fibronectins - biosynthesis Fibronectins - chemistry Fluorescent Antibody Technique, Indirect Glycosaminoglycans - metabolism Heparin - metabolism Immunoblotting Integrin alpha5beta1 - metabolism Mice Models, Genetic Peptides - chemistry Protein Binding Protein Isoforms Proteoglycans - metabolism Recombinant Proteins - chemistry RNA - metabolism RNA Splicing Transfection |
| title | Recombinant Expression and Characterization of a Novel Fibronectin Isoform Expressed in Cartilaginous Tissues |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-30T19%3A21%3A37IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Recombinant%20Expression%20and%20Characterization%20of%20a%20Novel%20Fibronectin%20Isoform%20Expressed%20in%20Cartilaginous%20Tissues&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Kozaki,%20Tomohiro&rft.date=2003-12-12&rft.volume=278&rft.issue=50&rft.spage=50546&rft.epage=50553&rft.pages=50546-50553&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M307432200&rft_dat=%3Cproquest_cross%3E19202767%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=19202767&rft_id=info:pmid/14525997&rft_els_id=S0021925820755766&rfr_iscdi=true |