Expression of biologically active rat apolipoprotein AIV in Escherichia coli
Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate–polyacrylamide gel elec...
Gespeichert in:
| Veröffentlicht in: | Physiology & behavior 2003, Vol.78 (1), p.149-155 |
|---|---|
| Hauptverfasser: | , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 155 |
|---|---|
| container_issue | 1 |
| container_start_page | 149 |
| container_title | Physiology & behavior |
| container_volume | 78 |
| creator | Liu, Min Maiorano, Nick Shen, Ling Pearson, Kevin Tajima, Daisuke Zhang, Dian Ming Woods, Stephen C Seeley, Randy J Davidson, W.Sean Tso, Patrick |
| description | Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in
Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis revealed that the isolated recombinant protein has a molecular mass of approximately 43 kDa, similar to that of natural rat apo AIV. Immunoblot analysis and N-terminal amino acid sequencing confirmed the identity of the recombinant apo AIV protein as natural rat apo AIV. The recombinant protein was functional in lipoprotein binding assays. Biological activity was assessed behaviorally in that the recombinant protein suppressed food intake of fasted rats comparably to natural rat apo AIV. Neither native nor recombinant apo AIV elicited a conditioned taste aversion (CTA) at doses that suppress feeding. These results indicate that the recombinant apo AIV is structurally and functionally indistinguishable from rat natural apo AIV, making this overexpression and purification scheme a powerful tool for future structure and function studies. |
| doi_str_mv | 10.1016/S0031-9384(02)00959-9 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_19769260</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0031938402009599</els_id><sourcerecordid>19769260</sourcerecordid><originalsourceid>FETCH-LOGICAL-c393t-f18886f4e795507fa5a35167f69cb7b048aaa33bc6989f8770520172077aef83</originalsourceid><addsrcrecordid>eNqFkE1rGzEQhkVpaBy3P6FhLy3JYdvRavV1KiE4H2DIoaH0JrTyKFFYrzbSOjT_vnJskmPmMgw8M_PyEPKVwg8KVPz8DcBorZlqT6A5BdBc1_oDmVElWc1B_v1IZq_IITnK-QFKsZZ9Ioe04UxA08zIcvFvTJhziEMVfdWF2Me74GzfP1fWTeEJq2Snyo6xD2McU5wwDNXZ9Z-qtEV295iCuw-2cgX4TA687TN-2fc5ub1Y3J5f1cuby-vzs2XtmGZT7alSSvgWpeYlqbfcMk6F9EK7TnbQKmstY50TWmmvpATeAJUNSGnRKzYn33dnS5zHDebJrEN22Pd2wLjJhmopdCOggHwHuhRzTujNmMLapmdDwWwtmheLZqvIQGNeLJZpTo73DzbdGldvW3ttBfi2B2wurnyygwv5jWs5bQVsk_7acVhsPAVMJruAg8NVSOgms4rhnSj_AZWpjbY</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>19769260</pqid></control><display><type>article</type><title>Expression of biologically active rat apolipoprotein AIV in Escherichia coli</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Liu, Min ; Maiorano, Nick ; Shen, Ling ; Pearson, Kevin ; Tajima, Daisuke ; Zhang, Dian Ming ; Woods, Stephen C ; Seeley, Randy J ; Davidson, W.Sean ; Tso, Patrick</creator><creatorcontrib>Liu, Min ; Maiorano, Nick ; Shen, Ling ; Pearson, Kevin ; Tajima, Daisuke ; Zhang, Dian Ming ; Woods, Stephen C ; Seeley, Randy J ; Davidson, W.Sean ; Tso, Patrick</creatorcontrib><description>Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in
Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis revealed that the isolated recombinant protein has a molecular mass of approximately 43 kDa, similar to that of natural rat apo AIV. Immunoblot analysis and N-terminal amino acid sequencing confirmed the identity of the recombinant apo AIV protein as natural rat apo AIV. The recombinant protein was functional in lipoprotein binding assays. Biological activity was assessed behaviorally in that the recombinant protein suppressed food intake of fasted rats comparably to natural rat apo AIV. Neither native nor recombinant apo AIV elicited a conditioned taste aversion (CTA) at doses that suppress feeding. These results indicate that the recombinant apo AIV is structurally and functionally indistinguishable from rat natural apo AIV, making this overexpression and purification scheme a powerful tool for future structure and function studies.</description><identifier>ISSN: 0031-9384</identifier><identifier>EISSN: 1873-507X</identifier><identifier>DOI: 10.1016/S0031-9384(02)00959-9</identifier><identifier>PMID: 12536022</identifier><language>eng</language><publisher>Cambridge: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animal ; Animals ; Apolipoprotein A-IV ; Apolipoproteins A - biosynthesis ; Apolipoproteins A - genetics ; Apolipoproteins A - pharmacology ; Avoidance Learning - drug effects ; Biological and medical sciences ; Biotechnology ; Body Weight - drug effects ; Conditioning ; DNA, Complementary - biosynthesis ; DNA, Complementary - genetics ; Eating - drug effects ; Escherichia coli ; Escherichia coli - metabolism ; Food intake ; Fundamental and applied biological sciences. Psychology ; Genetic engineering ; Genetic technics ; Immunoblotting ; Injections, Intraventricular ; Learning. Memory ; Lipoproteins - chemistry ; Lipoproteins - isolation & purification ; Male ; Methods. Procedures. Technologies ; Miscellaneous ; Molecular Sequence Data ; Psychology. Psychoanalysis. Psychiatry ; Psychology. Psychophysiology ; Rats ; Rats, Sprague-Dawley ; Recombinant expression ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - pharmacology ; Spectrometry, Mass, Electrospray Ionization ; Synthetic digonucleotides and genes. Sequencing ; Taste - drug effects</subject><ispartof>Physiology & behavior, 2003, Vol.78 (1), p.149-155</ispartof><rights>2003 Elsevier Science Inc.</rights><rights>2003 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c393t-f18886f4e795507fa5a35167f69cb7b048aaa33bc6989f8770520172077aef83</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0031-9384(02)00959-9$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,4024,27923,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14514608$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12536022$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Min</creatorcontrib><creatorcontrib>Maiorano, Nick</creatorcontrib><creatorcontrib>Shen, Ling</creatorcontrib><creatorcontrib>Pearson, Kevin</creatorcontrib><creatorcontrib>Tajima, Daisuke</creatorcontrib><creatorcontrib>Zhang, Dian Ming</creatorcontrib><creatorcontrib>Woods, Stephen C</creatorcontrib><creatorcontrib>Seeley, Randy J</creatorcontrib><creatorcontrib>Davidson, W.Sean</creatorcontrib><creatorcontrib>Tso, Patrick</creatorcontrib><title>Expression of biologically active rat apolipoprotein AIV in Escherichia coli</title><title>Physiology & behavior</title><addtitle>Physiol Behav</addtitle><description>Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in
Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis revealed that the isolated recombinant protein has a molecular mass of approximately 43 kDa, similar to that of natural rat apo AIV. Immunoblot analysis and N-terminal amino acid sequencing confirmed the identity of the recombinant apo AIV protein as natural rat apo AIV. The recombinant protein was functional in lipoprotein binding assays. Biological activity was assessed behaviorally in that the recombinant protein suppressed food intake of fasted rats comparably to natural rat apo AIV. Neither native nor recombinant apo AIV elicited a conditioned taste aversion (CTA) at doses that suppress feeding. These results indicate that the recombinant apo AIV is structurally and functionally indistinguishable from rat natural apo AIV, making this overexpression and purification scheme a powerful tool for future structure and function studies.</description><subject>Amino Acid Sequence</subject><subject>Animal</subject><subject>Animals</subject><subject>Apolipoprotein A-IV</subject><subject>Apolipoproteins A - biosynthesis</subject><subject>Apolipoproteins A - genetics</subject><subject>Apolipoproteins A - pharmacology</subject><subject>Avoidance Learning - drug effects</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Body Weight - drug effects</subject><subject>Conditioning</subject><subject>DNA, Complementary - biosynthesis</subject><subject>DNA, Complementary - genetics</subject><subject>Eating - drug effects</subject><subject>Escherichia coli</subject><subject>Escherichia coli - metabolism</subject><subject>Food intake</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Immunoblotting</subject><subject>Injections, Intraventricular</subject><subject>Learning. Memory</subject><subject>Lipoproteins - chemistry</subject><subject>Lipoproteins - isolation & purification</subject><subject>Male</subject><subject>Methods. Procedures. Technologies</subject><subject>Miscellaneous</subject><subject>Molecular Sequence Data</subject><subject>Psychology. Psychoanalysis. Psychiatry</subject><subject>Psychology. Psychophysiology</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Recombinant expression</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - pharmacology</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><subject>Synthetic digonucleotides and genes. Sequencing</subject><subject>Taste - drug effects</subject><issn>0031-9384</issn><issn>1873-507X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1rGzEQhkVpaBy3P6FhLy3JYdvRavV1KiE4H2DIoaH0JrTyKFFYrzbSOjT_vnJskmPmMgw8M_PyEPKVwg8KVPz8DcBorZlqT6A5BdBc1_oDmVElWc1B_v1IZq_IITnK-QFKsZZ9Ioe04UxA08zIcvFvTJhziEMVfdWF2Me74GzfP1fWTeEJq2Snyo6xD2McU5wwDNXZ9Z-qtEV295iCuw-2cgX4TA687TN-2fc5ub1Y3J5f1cuby-vzs2XtmGZT7alSSvgWpeYlqbfcMk6F9EK7TnbQKmstY50TWmmvpATeAJUNSGnRKzYn33dnS5zHDebJrEN22Pd2wLjJhmopdCOggHwHuhRzTujNmMLapmdDwWwtmheLZqvIQGNeLJZpTo73DzbdGldvW3ttBfi2B2wurnyygwv5jWs5bQVsk_7acVhsPAVMJruAg8NVSOgms4rhnSj_AZWpjbY</recordid><startdate>2003</startdate><enddate>2003</enddate><creator>Liu, Min</creator><creator>Maiorano, Nick</creator><creator>Shen, Ling</creator><creator>Pearson, Kevin</creator><creator>Tajima, Daisuke</creator><creator>Zhang, Dian Ming</creator><creator>Woods, Stephen C</creator><creator>Seeley, Randy J</creator><creator>Davidson, W.Sean</creator><creator>Tso, Patrick</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>7QG</scope><scope>7QL</scope><scope>7TK</scope><scope>C1K</scope></search><sort><creationdate>2003</creationdate><title>Expression of biologically active rat apolipoprotein AIV in Escherichia coli</title><author>Liu, Min ; Maiorano, Nick ; Shen, Ling ; Pearson, Kevin ; Tajima, Daisuke ; Zhang, Dian Ming ; Woods, Stephen C ; Seeley, Randy J ; Davidson, W.Sean ; Tso, Patrick</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c393t-f18886f4e795507fa5a35167f69cb7b048aaa33bc6989f8770520172077aef83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Amino Acid Sequence</topic><topic>Animal</topic><topic>Animals</topic><topic>Apolipoprotein A-IV</topic><topic>Apolipoproteins A - biosynthesis</topic><topic>Apolipoproteins A - genetics</topic><topic>Apolipoproteins A - pharmacology</topic><topic>Avoidance Learning - drug effects</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Body Weight - drug effects</topic><topic>Conditioning</topic><topic>DNA, Complementary - biosynthesis</topic><topic>DNA, Complementary - genetics</topic><topic>Eating - drug effects</topic><topic>Escherichia coli</topic><topic>Escherichia coli - metabolism</topic><topic>Food intake</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Immunoblotting</topic><topic>Injections, Intraventricular</topic><topic>Learning. Memory</topic><topic>Lipoproteins - chemistry</topic><topic>Lipoproteins - isolation & purification</topic><topic>Male</topic><topic>Methods. Procedures. Technologies</topic><topic>Miscellaneous</topic><topic>Molecular Sequence Data</topic><topic>Psychology. Psychoanalysis. Psychiatry</topic><topic>Psychology. Psychophysiology</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Recombinant expression</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - pharmacology</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><topic>Synthetic digonucleotides and genes. Sequencing</topic><topic>Taste - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Min</creatorcontrib><creatorcontrib>Maiorano, Nick</creatorcontrib><creatorcontrib>Shen, Ling</creatorcontrib><creatorcontrib>Pearson, Kevin</creatorcontrib><creatorcontrib>Tajima, Daisuke</creatorcontrib><creatorcontrib>Zhang, Dian Ming</creatorcontrib><creatorcontrib>Woods, Stephen C</creatorcontrib><creatorcontrib>Seeley, Randy J</creatorcontrib><creatorcontrib>Davidson, W.Sean</creatorcontrib><creatorcontrib>Tso, Patrick</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>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Neurosciences Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Physiology & behavior</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Min</au><au>Maiorano, Nick</au><au>Shen, Ling</au><au>Pearson, Kevin</au><au>Tajima, Daisuke</au><au>Zhang, Dian Ming</au><au>Woods, Stephen C</au><au>Seeley, Randy J</au><au>Davidson, W.Sean</au><au>Tso, Patrick</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of biologically active rat apolipoprotein AIV in Escherichia coli</atitle><jtitle>Physiology & behavior</jtitle><addtitle>Physiol Behav</addtitle><date>2003</date><risdate>2003</risdate><volume>78</volume><issue>1</issue><spage>149</spage><epage>155</epage><pages>149-155</pages><issn>0031-9384</issn><eissn>1873-507X</eissn><abstract>Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in
Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis revealed that the isolated recombinant protein has a molecular mass of approximately 43 kDa, similar to that of natural rat apo AIV. Immunoblot analysis and N-terminal amino acid sequencing confirmed the identity of the recombinant apo AIV protein as natural rat apo AIV. The recombinant protein was functional in lipoprotein binding assays. Biological activity was assessed behaviorally in that the recombinant protein suppressed food intake of fasted rats comparably to natural rat apo AIV. Neither native nor recombinant apo AIV elicited a conditioned taste aversion (CTA) at doses that suppress feeding. These results indicate that the recombinant apo AIV is structurally and functionally indistinguishable from rat natural apo AIV, making this overexpression and purification scheme a powerful tool for future structure and function studies.</abstract><cop>Cambridge</cop><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>12536022</pmid><doi>10.1016/S0031-9384(02)00959-9</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0031-9384 |
| ispartof | Physiology & behavior, 2003, Vol.78 (1), p.149-155 |
| issn | 0031-9384 1873-507X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_19769260 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Amino Acid Sequence Animal Animals Apolipoprotein A-IV Apolipoproteins A - biosynthesis Apolipoproteins A - genetics Apolipoproteins A - pharmacology Avoidance Learning - drug effects Biological and medical sciences Biotechnology Body Weight - drug effects Conditioning DNA, Complementary - biosynthesis DNA, Complementary - genetics Eating - drug effects Escherichia coli Escherichia coli - metabolism Food intake Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Immunoblotting Injections, Intraventricular Learning. Memory Lipoproteins - chemistry Lipoproteins - isolation & purification Male Methods. Procedures. Technologies Miscellaneous Molecular Sequence Data Psychology. Psychoanalysis. Psychiatry Psychology. Psychophysiology Rats Rats, Sprague-Dawley Recombinant expression Recombinant Proteins - biosynthesis Recombinant Proteins - pharmacology Spectrometry, Mass, Electrospray Ionization Synthetic digonucleotides and genes. Sequencing Taste - drug effects |
| title | Expression of biologically active rat apolipoprotein AIV in Escherichia coli |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-03T17%3A18%3A33IST&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=Expression%20of%20biologically%20active%20rat%20apolipoprotein%20AIV%20in%20Escherichia%20coli&rft.jtitle=Physiology%20&%20behavior&rft.au=Liu,%20Min&rft.date=2003&rft.volume=78&rft.issue=1&rft.spage=149&rft.epage=155&rft.pages=149-155&rft.issn=0031-9384&rft.eissn=1873-507X&rft_id=info:doi/10.1016/S0031-9384(02)00959-9&rft_dat=%3Cproquest_cross%3E19769260%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=19769260&rft_id=info:pmid/12536022&rft_els_id=S0031938402009599&rfr_iscdi=true |