Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector
The Ku autoantigen is a human nuclear, DNA-binding heterodimer of 70kDa and 86kDa proteins. It is the target of autoantibodies in several autoimmune diseases. We now report the expression of a cDNA encoding the 70kDa Ku protein. Large amounts of protein were obtained using a recombinant baculovirus...
Gespeichert in:
| Veröffentlicht in: | Biochemical and biophysical research communications 1990-04, Vol.168 (2), p.747-755 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 755 |
|---|---|
| container_issue | 2 |
| container_start_page | 747 |
| container_title | Biochemical and biophysical research communications |
| container_volume | 168 |
| creator | Allaway, Graham P. Vivino, Alfred A. Kohn, Leonard D. Notkins, Abner Louis Bellur, S. Prabhakar |
| description | The Ku autoantigen is a human nuclear, DNA-binding heterodimer of 70kDa and 86kDa proteins. It is the target of autoantibodies in several autoimmune diseases. We now report the expression of a cDNA encoding the 70kDa Ku protein. Large amounts of protein were obtained using a recombinant baculovirus vector, in contrast with earlier unsuccessful attempts using other expression systems. We demonstrate that the 70kDa Ku protein is targeted to the nucleus and is associated with the nuclear matrix when expressed in the absence of the 86kDa Ku component. No post-translational modifications were observed. The 70kDa protein binds double and single-stranded DNA with very high affinity. Our results suggest that the baculovirus expression system may be of widespread use in the production and characterization of human autoantigens. |
| doi_str_mv | 10.1016/0006-291X(90)92385-D |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_79760063</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>0006291X9092385D</els_id><sourcerecordid>79760063</sourcerecordid><originalsourceid>FETCH-LOGICAL-c387t-7bc44615ea41d99f05dee2e5315afa8682960bf1864d19529450ca4cc958a6593</originalsourceid><addsrcrecordid>eNp9kU2LFDEQhoMo6-zqP1DIRVkPrUl3Ot25CDLjFy54UfAWqtPVO9HuZDYfg_oT_NVmnNG9CYGC1PMWxVOEPOLsOWdcvmCMyapW_MulYs9U3fRttblDVpwpVtWcibtk9Q-5T85j_MoY50KqM3LGpaprLlfk13oLAUzCYH9Cst5RP9G0Rdqxbxugxi8779Clv9_bvICjHzKFnDy4ZK_RUfy-CxgjjtS68iKaRA3OM3XZzGhpjtZdU6ABy7zBupKjA5g8-70NOdJ9CfjwgNybYI748FQvyOc3rz-t31VXH9--X7-6qkzTd6nqBiOE5C2C4KNSE2tHxBrbhrcwQS_7Wkk2TLyXYuSqrZVomQFhjGp7kK1qLsjT49xd8DcZY9KLjYd1waHPUXeqk8VbU0BxBE3wMQac9C7YBcIPzZk-nEAf_OqDX62Y_nMCvSmxx6f5eVhwvA0dnZf-k1MfooF5CuCMjbeYEr1QNS_cyyOHRcbeYtDRWHQGR1tEJj16-_9FfgPDOKS4</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>79760063</pqid></control><display><type>article</type><title>Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector</title><source>MEDLINE</source><source>ScienceDirect Journals (5 years ago - present)</source><creator>Allaway, Graham P. ; Vivino, Alfred A. ; Kohn, Leonard D. ; Notkins, Abner Louis ; Bellur, S. Prabhakar</creator><creatorcontrib>Allaway, Graham P. ; Vivino, Alfred A. ; Kohn, Leonard D. ; Notkins, Abner Louis ; Bellur, S. Prabhakar</creatorcontrib><description>The Ku autoantigen is a human nuclear, DNA-binding heterodimer of 70kDa and 86kDa proteins. It is the target of autoantibodies in several autoimmune diseases. We now report the expression of a cDNA encoding the 70kDa Ku protein. Large amounts of protein were obtained using a recombinant baculovirus vector, in contrast with earlier unsuccessful attempts using other expression systems. We demonstrate that the 70kDa Ku protein is targeted to the nucleus and is associated with the nuclear matrix when expressed in the absence of the 86kDa Ku component. No post-translational modifications were observed. The 70kDa protein binds double and single-stranded DNA with very high affinity. Our results suggest that the baculovirus expression system may be of widespread use in the production and characterization of human autoantigens.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/0006-291X(90)92385-D</identifier><identifier>PMID: 1692216</identifier><identifier>CODEN: BBRCA9</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Animals ; Antigens, Nuclear ; Antigens, Surface - genetics ; Autoantigens - genetics ; Biological and medical sciences ; Cell Nucleus - metabolism ; DNA - analysis ; DNA Helicases ; DNA-Binding Proteins - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetic Vectors ; Humans ; In Vitro Techniques ; Ku Autoantigen ; Lepidoptera - genetics ; Lepidoptera - immunology ; Molecular and cellular biology ; Molecular genetics ; Molecular Weight ; Protein Processing, Post-Translational ; Recombination, Genetic ; RNA - analysis ; Transfection</subject><ispartof>Biochemical and biophysical research communications, 1990-04, Vol.168 (2), p.747-755</ispartof><rights>1990 Academic Press, Inc.</rights><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c387t-7bc44615ea41d99f05dee2e5315afa8682960bf1864d19529450ca4cc958a6593</citedby><cites>FETCH-LOGICAL-c387t-7bc44615ea41d99f05dee2e5315afa8682960bf1864d19529450ca4cc958a6593</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0006-291X(90)92385-D$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19484921$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1692216$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Allaway, Graham P.</creatorcontrib><creatorcontrib>Vivino, Alfred A.</creatorcontrib><creatorcontrib>Kohn, Leonard D.</creatorcontrib><creatorcontrib>Notkins, Abner Louis</creatorcontrib><creatorcontrib>Bellur, S. Prabhakar</creatorcontrib><title>Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>The Ku autoantigen is a human nuclear, DNA-binding heterodimer of 70kDa and 86kDa proteins. It is the target of autoantibodies in several autoimmune diseases. We now report the expression of a cDNA encoding the 70kDa Ku protein. Large amounts of protein were obtained using a recombinant baculovirus vector, in contrast with earlier unsuccessful attempts using other expression systems. We demonstrate that the 70kDa Ku protein is targeted to the nucleus and is associated with the nuclear matrix when expressed in the absence of the 86kDa Ku component. No post-translational modifications were observed. The 70kDa protein binds double and single-stranded DNA with very high affinity. Our results suggest that the baculovirus expression system may be of widespread use in the production and characterization of human autoantigens.</description><subject>Animals</subject><subject>Antigens, Nuclear</subject><subject>Antigens, Surface - genetics</subject><subject>Autoantigens - genetics</subject><subject>Biological and medical sciences</subject><subject>Cell Nucleus - metabolism</subject><subject>DNA - analysis</subject><subject>DNA Helicases</subject><subject>DNA-Binding Proteins - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic Vectors</subject><subject>Humans</subject><subject>In Vitro Techniques</subject><subject>Ku Autoantigen</subject><subject>Lepidoptera - genetics</subject><subject>Lepidoptera - immunology</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Weight</subject><subject>Protein Processing, Post-Translational</subject><subject>Recombination, Genetic</subject><subject>RNA - analysis</subject><subject>Transfection</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU2LFDEQhoMo6-zqP1DIRVkPrUl3Ot25CDLjFy54UfAWqtPVO9HuZDYfg_oT_NVmnNG9CYGC1PMWxVOEPOLsOWdcvmCMyapW_MulYs9U3fRttblDVpwpVtWcibtk9Q-5T85j_MoY50KqM3LGpaprLlfk13oLAUzCYH9Cst5RP9G0Rdqxbxugxi8779Clv9_bvICjHzKFnDy4ZK_RUfy-CxgjjtS68iKaRA3OM3XZzGhpjtZdU6ABy7zBupKjA5g8-70NOdJ9CfjwgNybYI748FQvyOc3rz-t31VXH9--X7-6qkzTd6nqBiOE5C2C4KNSE2tHxBrbhrcwQS_7Wkk2TLyXYuSqrZVomQFhjGp7kK1qLsjT49xd8DcZY9KLjYd1waHPUXeqk8VbU0BxBE3wMQac9C7YBcIPzZk-nEAf_OqDX62Y_nMCvSmxx6f5eVhwvA0dnZf-k1MfooF5CuCMjbeYEr1QNS_cyyOHRcbeYtDRWHQGR1tEJj16-_9FfgPDOKS4</recordid><startdate>19900430</startdate><enddate>19900430</enddate><creator>Allaway, Graham P.</creator><creator>Vivino, Alfred A.</creator><creator>Kohn, Leonard D.</creator><creator>Notkins, Abner Louis</creator><creator>Bellur, S. Prabhakar</creator><general>Elsevier Inc</general><general>Elsevier</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>7X8</scope></search><sort><creationdate>19900430</creationdate><title>Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector</title><author>Allaway, Graham P. ; Vivino, Alfred A. ; Kohn, Leonard D. ; Notkins, Abner Louis ; Bellur, S. Prabhakar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c387t-7bc44615ea41d99f05dee2e5315afa8682960bf1864d19529450ca4cc958a6593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Animals</topic><topic>Antigens, Nuclear</topic><topic>Antigens, Surface - genetics</topic><topic>Autoantigens - genetics</topic><topic>Biological and medical sciences</topic><topic>Cell Nucleus - metabolism</topic><topic>DNA - analysis</topic><topic>DNA Helicases</topic><topic>DNA-Binding Proteins - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic Vectors</topic><topic>Humans</topic><topic>In Vitro Techniques</topic><topic>Ku Autoantigen</topic><topic>Lepidoptera - genetics</topic><topic>Lepidoptera - immunology</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Weight</topic><topic>Protein Processing, Post-Translational</topic><topic>Recombination, Genetic</topic><topic>RNA - analysis</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Allaway, Graham P.</creatorcontrib><creatorcontrib>Vivino, Alfred A.</creatorcontrib><creatorcontrib>Kohn, Leonard D.</creatorcontrib><creatorcontrib>Notkins, Abner Louis</creatorcontrib><creatorcontrib>Bellur, S. Prabhakar</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>MEDLINE - Academic</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Allaway, Graham P.</au><au>Vivino, Alfred A.</au><au>Kohn, Leonard D.</au><au>Notkins, Abner Louis</au><au>Bellur, S. Prabhakar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>1990-04-30</date><risdate>1990</risdate><volume>168</volume><issue>2</issue><spage>747</spage><epage>755</epage><pages>747-755</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><coden>BBRCA9</coden><abstract>The Ku autoantigen is a human nuclear, DNA-binding heterodimer of 70kDa and 86kDa proteins. It is the target of autoantibodies in several autoimmune diseases. We now report the expression of a cDNA encoding the 70kDa Ku protein. Large amounts of protein were obtained using a recombinant baculovirus vector, in contrast with earlier unsuccessful attempts using other expression systems. We demonstrate that the 70kDa Ku protein is targeted to the nucleus and is associated with the nuclear matrix when expressed in the absence of the 86kDa Ku component. No post-translational modifications were observed. The 70kDa protein binds double and single-stranded DNA with very high affinity. Our results suggest that the baculovirus expression system may be of widespread use in the production and characterization of human autoantigens.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>1692216</pmid><doi>10.1016/0006-291X(90)92385-D</doi><tpages>9</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0006-291X |
| ispartof | Biochemical and biophysical research communications, 1990-04, Vol.168 (2), p.747-755 |
| issn | 0006-291X 1090-2104 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79760063 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Animals Antigens, Nuclear Antigens, Surface - genetics Autoantigens - genetics Biological and medical sciences Cell Nucleus - metabolism DNA - analysis DNA Helicases DNA-Binding Proteins - genetics Fundamental and applied biological sciences. Psychology Gene Expression Genetic Vectors Humans In Vitro Techniques Ku Autoantigen Lepidoptera - genetics Lepidoptera - immunology Molecular and cellular biology Molecular genetics Molecular Weight Protein Processing, Post-Translational Recombination, Genetic RNA - analysis Transfection |
| title | Characterization of the 70kDa component of the human Ku autoantigen expressed in insect cell nuclei using a recombinant baculovirus vector |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-12T11%3A53%3A48IST&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=Characterization%20of%20the%2070kDa%20component%20of%20the%20human%20Ku%20autoantigen%20expressed%20in%20insect%20cell%20nuclei%20using%20a%20recombinant%20baculovirus%20vector&rft.jtitle=Biochemical%20and%20biophysical%20research%20communications&rft.au=Allaway,%20Graham%20P.&rft.date=1990-04-30&rft.volume=168&rft.issue=2&rft.spage=747&rft.epage=755&rft.pages=747-755&rft.issn=0006-291X&rft.eissn=1090-2104&rft.coden=BBRCA9&rft_id=info:doi/10.1016/0006-291X(90)92385-D&rft_dat=%3Cproquest_cross%3E79760063%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=79760063&rft_id=info:pmid/1692216&rft_els_id=0006291X9092385D&rfr_iscdi=true |