Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β
Expression plasmids carrying the coding sequence of mature human interleukin 1β(ILIβ) linked either to a Met start codon, or fused to different efficient Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membr...
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| Veröffentlicht in: | Gene 1989-12, Vol.85 (2), p.499-510 |
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| creator | Denèfle, Patrice Kovarik, Sylvie Ciora, Thierry Gosselet, Nicole Bénichou, Jean-Claude Latta, Martine Guinet, Françoise Ryter, Antoinette Mayaux, Jean-François |
| description | Expression plasmids carrying the coding sequence of mature human interleukin 1β(ILIβ) linked either to a Met start codon, or fused to different efficient
Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membrane protein (OmpA) or from a periplasmic protein (PhoA). The synthesis of IL1β from these fusions was investigated in an otherwise strictly isogenic context using identical conditions of derepression and culture media. The Met-IL1β fusion produced a soluble cytoplasmic protein which could be released from the cells by osmotic shock whereas the OmpA and PhoA fusions were always insoluble. The extent of
sOmpA-IL1βmaturation was found to vary from 50 to 100%, mainly depending on the medium used, whereas no significant maturation of the signal peptide could be detected in the case of the
sPhoA-IL1β fusion. Immuno-electron microscopy revealed that the
sOmpA-IL 1β fusion was targeted to the inner membrane, whereas the ThoA-IL 1βfusion remained within the cytoplasm and thus did not appear to enter the secretion pathway. Amplifying the
E. coli signal peptidase
lep gene on a multicopy plasmid did not improve signal peptide removal from
sOmpA-IL1β. Moreover, these
E. coli |
| doi_str_mv | 10.1016/0378-1119(89)90444-7 |
| format | Article |
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Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membrane protein (OmpA) or from a periplasmic protein (PhoA). The synthesis of IL1β from these fusions was investigated in an otherwise strictly isogenic context using identical conditions of derepression and culture media. The Met-IL1β fusion produced a soluble cytoplasmic protein which could be released from the cells by osmotic shock whereas the OmpA and PhoA fusions were always insoluble. The extent of
sOmpA-IL1βmaturation was found to vary from 50 to 100%, mainly depending on the medium used, whereas no significant maturation of the signal peptide could be detected in the case of the
sPhoA-IL1β fusion. Immuno-electron microscopy revealed that the
sOmpA-IL 1β fusion was targeted to the inner membrane, whereas the ThoA-IL 1βfusion remained within the cytoplasm and thus did not appear to enter the secretion pathway. Amplifying the
E. coli signal peptidase
lep gene on a multicopy plasmid did not improve signal peptide removal from
sOmpA-IL1β. Moreover, these
E. coli</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(89)90444-7</identifier><identifier>PMID: 2697645</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Amino Acid Sequence ; Antiviral drugs ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli - genetics ; Escherichia coli - ultrastructure ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetic engineering ; Genetic technics ; Genetic Vectors ; Health. Pharmaceutical industry ; Humans ; immuno-electron microscopy ; Industrial applications and implications. Economical aspects ; Interleukin-1 - biosynthesis ; Interleukin-1 - genetics ; Interleukin-1 - metabolism ; Methods. Procedures. Technologies ; Molecular Sequence Data ; Molecular Weight ; Oligonucleotide Probes ; periplasm ; plasmid vectors ; Plasmids ; Production of active biomolecules ; protein aggregation ; Protein Sorting Signals - metabolism ; protein translocation ; Recombinant DNA ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - metabolism ; Restriction Mapping ; signal peptide removal</subject><ispartof>Gene, 1989-12, Vol.85 (2), p.499-510</ispartof><rights>1989</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-47dc0eb7d9e7240d0cda0987d9c197ef93648110535d2ed101d739bff8a5b4fe3</citedby><cites>FETCH-LOGICAL-c422t-47dc0eb7d9e7240d0cda0987d9c197ef93648110535d2ed101d739bff8a5b4fe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0378-1119(89)90444-7$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6894252$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2697645$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Denèfle, Patrice</creatorcontrib><creatorcontrib>Kovarik, Sylvie</creatorcontrib><creatorcontrib>Ciora, Thierry</creatorcontrib><creatorcontrib>Gosselet, Nicole</creatorcontrib><creatorcontrib>Bénichou, Jean-Claude</creatorcontrib><creatorcontrib>Latta, Martine</creatorcontrib><creatorcontrib>Guinet, Françoise</creatorcontrib><creatorcontrib>Ryter, Antoinette</creatorcontrib><creatorcontrib>Mayaux, Jean-François</creatorcontrib><title>Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β</title><title>Gene</title><addtitle>Gene</addtitle><description>Expression plasmids carrying the coding sequence of mature human interleukin 1β(ILIβ) linked either to a Met start codon, or fused to different efficient
Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membrane protein (OmpA) or from a periplasmic protein (PhoA). The synthesis of IL1β from these fusions was investigated in an otherwise strictly isogenic context using identical conditions of derepression and culture media. The Met-IL1β fusion produced a soluble cytoplasmic protein which could be released from the cells by osmotic shock whereas the OmpA and PhoA fusions were always insoluble. The extent of
sOmpA-IL1βmaturation was found to vary from 50 to 100%, mainly depending on the medium used, whereas no significant maturation of the signal peptide could be detected in the case of the
sPhoA-IL1β fusion. Immuno-electron microscopy revealed that the
sOmpA-IL 1β fusion was targeted to the inner membrane, whereas the ThoA-IL 1βfusion remained within the cytoplasm and thus did not appear to enter the secretion pathway. Amplifying the
E. coli signal peptidase
lep gene on a multicopy plasmid did not improve signal peptide removal from
sOmpA-IL1β. Moreover, these
E. coli</description><subject>Amino Acid Sequence</subject><subject>Antiviral drugs</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning, Molecular</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - ultrastructure</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Vectors</subject><subject>Health. Pharmaceutical industry</subject><subject>Humans</subject><subject>immuno-electron microscopy</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Interleukin-1 - biosynthesis</subject><subject>Interleukin-1 - genetics</subject><subject>Interleukin-1 - metabolism</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Oligonucleotide Probes</subject><subject>periplasm</subject><subject>plasmid vectors</subject><subject>Plasmids</subject><subject>Production of active biomolecules</subject><subject>protein aggregation</subject><subject>Protein Sorting Signals - metabolism</subject><subject>protein translocation</subject><subject>Recombinant DNA</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - metabolism</subject><subject>Restriction Mapping</subject><subject>signal peptide removal</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM-OFCEQh4nRrOPqG2jCwRg9tAJNN80eTMxmdU028aJnQkOxgzJNC91GLz6UD-IzWeOMc5RL8eerX6iPkMecveSM969Yq4aGc66fD_qFZlLKRt0hGz4o3TDWDnfJ5oTcJw9q_cxwdZ04I2ei16qX3Yb8vIYFSk75Nq-VziUvECcK3-dcFoq7q-q2UKLbRktdTvECL0NaYXJAc6CjddgebaI13k5YZpiX6KHSPNFlC_-SEN2uOzthN_IJ1i-YzX__ekjuBZsqPDrWc_Lp7dXHy-vm5sO795dvbhonhVgaqbxjMCqvQQnJPHPeMj3g2XGtIOi2lwPnrGs7L8CjHq9aPYYw2G6UAdpz8uyQixN-XaEuZherg5TsBDi4UVpqgYkIygPoSq61QDBziTtbfhjOzF672Ts1e6dm0OavdqOw7ckxfx134E9NR8_4_vT4bquzKRQ7uVhPWD9oKTqB2OsDBujiW4Riqot71z4WcIvxOf7_H38A-06hKw</recordid><startdate>19891228</startdate><enddate>19891228</enddate><creator>Denèfle, Patrice</creator><creator>Kovarik, Sylvie</creator><creator>Ciora, Thierry</creator><creator>Gosselet, Nicole</creator><creator>Bénichou, Jean-Claude</creator><creator>Latta, Martine</creator><creator>Guinet, Françoise</creator><creator>Ryter, Antoinette</creator><creator>Mayaux, Jean-François</creator><general>Elsevier B.V</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>19891228</creationdate><title>Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β</title><author>Denèfle, Patrice ; Kovarik, Sylvie ; Ciora, Thierry ; Gosselet, Nicole ; Bénichou, Jean-Claude ; Latta, Martine ; Guinet, Françoise ; Ryter, Antoinette ; Mayaux, Jean-François</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-47dc0eb7d9e7240d0cda0987d9c197ef93648110535d2ed101d739bff8a5b4fe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Amino Acid Sequence</topic><topic>Antiviral drugs</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cloning, Molecular</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - ultrastructure</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genetic Vectors</topic><topic>Health. Pharmaceutical industry</topic><topic>Humans</topic><topic>immuno-electron microscopy</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>Interleukin-1 - biosynthesis</topic><topic>Interleukin-1 - genetics</topic><topic>Interleukin-1 - metabolism</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Oligonucleotide Probes</topic><topic>periplasm</topic><topic>plasmid vectors</topic><topic>Plasmids</topic><topic>Production of active biomolecules</topic><topic>protein aggregation</topic><topic>Protein Sorting Signals - metabolism</topic><topic>protein translocation</topic><topic>Recombinant DNA</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - metabolism</topic><topic>Restriction Mapping</topic><topic>signal peptide removal</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Denèfle, Patrice</creatorcontrib><creatorcontrib>Kovarik, Sylvie</creatorcontrib><creatorcontrib>Ciora, Thierry</creatorcontrib><creatorcontrib>Gosselet, Nicole</creatorcontrib><creatorcontrib>Bénichou, Jean-Claude</creatorcontrib><creatorcontrib>Latta, Martine</creatorcontrib><creatorcontrib>Guinet, Françoise</creatorcontrib><creatorcontrib>Ryter, Antoinette</creatorcontrib><creatorcontrib>Mayaux, Jean-François</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>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Denèfle, Patrice</au><au>Kovarik, Sylvie</au><au>Ciora, Thierry</au><au>Gosselet, Nicole</au><au>Bénichou, Jean-Claude</au><au>Latta, Martine</au><au>Guinet, Françoise</au><au>Ryter, Antoinette</au><au>Mayaux, Jean-François</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1989-12-28</date><risdate>1989</risdate><volume>85</volume><issue>2</issue><spage>499</spage><epage>510</epage><pages>499-510</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>Expression plasmids carrying the coding sequence of mature human interleukin 1β(ILIβ) linked either to a Met start codon, or fused to different efficient
Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membrane protein (OmpA) or from a periplasmic protein (PhoA). The synthesis of IL1β from these fusions was investigated in an otherwise strictly isogenic context using identical conditions of derepression and culture media. The Met-IL1β fusion produced a soluble cytoplasmic protein which could be released from the cells by osmotic shock whereas the OmpA and PhoA fusions were always insoluble. The extent of
sOmpA-IL1βmaturation was found to vary from 50 to 100%, mainly depending on the medium used, whereas no significant maturation of the signal peptide could be detected in the case of the
sPhoA-IL1β fusion. Immuno-electron microscopy revealed that the
sOmpA-IL 1β fusion was targeted to the inner membrane, whereas the ThoA-IL 1βfusion remained within the cytoplasm and thus did not appear to enter the secretion pathway. Amplifying the
E. coli signal peptidase
lep gene on a multicopy plasmid did not improve signal peptide removal from
sOmpA-IL1β. Moreover, these
E. coli</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>2697645</pmid><doi>10.1016/0378-1119(89)90444-7</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1119 |
| ispartof | Gene, 1989-12, Vol.85 (2), p.499-510 |
| issn | 0378-1119 1879-0038 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_79492098 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Amino Acid Sequence Antiviral drugs Base Sequence Biological and medical sciences Biotechnology Cloning, Molecular Electrophoresis, Polyacrylamide Gel Escherichia coli - genetics Escherichia coli - ultrastructure Fundamental and applied biological sciences. Psychology Gene Expression Genetic engineering Genetic technics Genetic Vectors Health. Pharmaceutical industry Humans immuno-electron microscopy Industrial applications and implications. Economical aspects Interleukin-1 - biosynthesis Interleukin-1 - genetics Interleukin-1 - metabolism Methods. Procedures. Technologies Molecular Sequence Data Molecular Weight Oligonucleotide Probes periplasm plasmid vectors Plasmids Production of active biomolecules protein aggregation Protein Sorting Signals - metabolism protein translocation Recombinant DNA Recombinant Proteins - biosynthesis Recombinant Proteins - metabolism Restriction Mapping signal peptide removal |
| title | Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β |
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