Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli
cDNA clones encoding proteins of approximately 18 kDa in which 83% of the amino acids are conserved relative to the published sequences of mammalian cyclophilin/rotamase (CyP) have been isolated from tomato, maize, and Brassica napus. In correspondence with the mammalian genes, but in contrast with...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1990-12, Vol.87 (24), p.9519-9523 |
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| creator | GASSER, C. S GUNNING, D. A BUDELIER, K. A BROWN, S. M |
| description | cDNA clones encoding proteins of approximately 18 kDa in which 83% of the amino acids are conserved relative to the published sequences of mammalian cyclophilin/rotamase (CyP) have been isolated from tomato, maize, and Brassica napus. In correspondence with the mammalian genes, but in contrast with the Neurospora gene and one yeast CyP gene, the plant CyP genes encode only mature proteins lacking transit peptides. RNA blot analyses demonstrate that CyP genes are expressed in all plant organs tested. Southern blots of genomic DNA indicate that there are small families (two to eight members) of CyP-related genes in maize and B. napus. A vector was constructed for expression of the tomato cDNA in E. coli. SDS/polyacrylamide gels show that extracts of appropriately induced cells harboring this vector contain nearly 40% of the protein as a single approximately 18-kDa band. While the majority of this protein is sequestered in insoluble inclusion bodies, the soluble extracts have higher levels of peptidyl-prolyl cis-trans isomerase (rotamase) activity than extracts of wild-type cells. This additional activity is sensitive to inhibition by the cyclic undecapeptide cyclosporin A. |
| doi_str_mv | 10.1073/pnas.87.24.9519 |
| format | Article |
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S ; GUNNING, D. A ; BUDELIER, K. A ; BROWN, S. M</creator><creatorcontrib>GASSER, C. S ; GUNNING, D. A ; BUDELIER, K. A ; BROWN, S. M</creatorcontrib><description>cDNA clones encoding proteins of approximately 18 kDa in which 83% of the amino acids are conserved relative to the published sequences of mammalian cyclophilin/rotamase (CyP) have been isolated from tomato, maize, and Brassica napus. In correspondence with the mammalian genes, but in contrast with the Neurospora gene and one yeast CyP gene, the plant CyP genes encode only mature proteins lacking transit peptides. RNA blot analyses demonstrate that CyP genes are expressed in all plant organs tested. Southern blots of genomic DNA indicate that there are small families (two to eight members) of CyP-related genes in maize and B. napus. A vector was constructed for expression of the tomato cDNA in E. coli. SDS/polyacrylamide gels show that extracts of appropriately induced cells harboring this vector contain nearly 40% of the protein as a single approximately 18-kDa band. While the majority of this protein is sequestered in insoluble inclusion bodies, the soluble extracts have higher levels of peptidyl-prolyl cis-trans isomerase (rotamase) activity than extracts of wild-type cells. This additional activity is sensitive to inhibition by the cyclic undecapeptide cyclosporin A.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.87.24.9519</identifier><identifier>PMID: 1702215</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Acad Sciences</publisher><subject>Amino Acid Isomerases - genetics ; Amino Acid Isomerases - isolation & purification ; Amino Acid Isomerases - metabolism ; Amino Acid Sequence ; Base Sequence ; Biological and medical sciences ; Blotting, Southern ; Carrier Proteins - genetics ; Carrier Proteins - isolation & purification ; Carrier Proteins - metabolism ; cDNA ; Cloning, Molecular ; cyclophilin ; Cytosol - enzymology ; DNA - genetics ; DNA - isolation & purification ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Library ; genes ; Genes. Genome ; Kinetics ; Lycopersicon esculentum ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Molecular Weight ; nucleotide sequence ; Oligonucleotide Probes ; Peptidylprolyl Isomerase ; Plants - enzymology ; Plants - genetics ; predictions ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; RNA - genetics ; RNA - isolation & purification</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1990-12, Vol.87 (24), p.9519-9523</ispartof><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c434t-60914f858de6a0a8a24cc30ffae569b86d099aadb790287b93ca1427b86c0b583</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/87/24.cover.gif</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC55203/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC55203/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27923,27924,53790,53792</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19602312$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1702215$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>GASSER, C. S</creatorcontrib><creatorcontrib>GUNNING, D. A</creatorcontrib><creatorcontrib>BUDELIER, K. A</creatorcontrib><creatorcontrib>BROWN, S. M</creatorcontrib><title>Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>cDNA clones encoding proteins of approximately 18 kDa in which 83% of the amino acids are conserved relative to the published sequences of mammalian cyclophilin/rotamase (CyP) have been isolated from tomato, maize, and Brassica napus. In correspondence with the mammalian genes, but in contrast with the Neurospora gene and one yeast CyP gene, the plant CyP genes encode only mature proteins lacking transit peptides. RNA blot analyses demonstrate that CyP genes are expressed in all plant organs tested. Southern blots of genomic DNA indicate that there are small families (two to eight members) of CyP-related genes in maize and B. napus. A vector was constructed for expression of the tomato cDNA in E. coli. SDS/polyacrylamide gels show that extracts of appropriately induced cells harboring this vector contain nearly 40% of the protein as a single approximately 18-kDa band. While the majority of this protein is sequestered in insoluble inclusion bodies, the soluble extracts have higher levels of peptidyl-prolyl cis-trans isomerase (rotamase) activity than extracts of wild-type cells. This additional activity is sensitive to inhibition by the cyclic undecapeptide cyclosporin A.</description><subject>Amino Acid Isomerases - genetics</subject><subject>Amino Acid Isomerases - isolation & purification</subject><subject>Amino Acid Isomerases - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Carrier Proteins - genetics</subject><subject>Carrier Proteins - isolation & purification</subject><subject>Carrier Proteins - metabolism</subject><subject>cDNA</subject><subject>Cloning, Molecular</subject><subject>cyclophilin</subject><subject>Cytosol - enzymology</subject><subject>DNA - genetics</subject><subject>DNA - isolation & purification</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Library</subject><subject>genes</subject><subject>Genes. Genome</subject><subject>Kinetics</subject><subject>Lycopersicon esculentum</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>nucleotide sequence</subject><subject>Oligonucleotide Probes</subject><subject>Peptidylprolyl Isomerase</subject><subject>Plants - enzymology</subject><subject>Plants - genetics</subject><subject>predictions</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>RNA - genetics</subject><subject>RNA - isolation & purification</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkk2P1SAUhonRjNfRtStNN-qq9wKFFpLZmMn4kUziQl2TU0qnGG6pQCdz_5C_U-qt3nGjCQkk7_OeD85B6DnBW4KbajeNELei2VK2lZzIB2hDsCRlzSR-iDYY06YUjLLH6EmM3zDGkgt8hs5IgyklfIN-fE5h1mkOpoCxK8zdFEyM1o-F7wt9SD56Z3V-aeenwTo77iYzJdsdXDkF7w6u0DaWKcAYCxv93gSIZjEP9mYwoZgcjCn-Cp75Ludag0N-3Zoi-T0kfz9Bkc9V1Nls9WCh0LmCp-hRDy6aZ-t9jr6-u_py-aG8_vT-4-Xb61KziqWyzr2zXnDRmRowCKBM6wr3PRhey1bUHZYSoGsbialoWllpIIw2WdG45aI6RxfHuNPc7k2nzZg7c2oKdg_hoDxY9bcy2kHd-FvFOcVVtr9e7cF_n01Mam-jNi7_gfFzVAITQQnh_wVJzYWUTGZwdwR18DEG0_-phWC1bIBaNkCJRlGmlg3Ijhf3Wzjxx5Fn_dWqQ9Tg-jy5PMETJmtMK0Iz92bllgS_5VMi1c_OJXOXMvnyn2T1Ey5Z2C4</recordid><startdate>19901201</startdate><enddate>19901201</enddate><creator>GASSER, C. S</creator><creator>GUNNING, D. A</creator><creator>BUDELIER, K. A</creator><creator>BROWN, S. M</creator><general>National Acad Sciences</general><general>National Academy of Sciences of the United States of America</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>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M81</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19901201</creationdate><title>Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli</title><author>GASSER, C. S ; GUNNING, D. A ; BUDELIER, K. A ; BROWN, S. M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c434t-60914f858de6a0a8a24cc30ffae569b86d099aadb790287b93ca1427b86c0b583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Amino Acid Isomerases - genetics</topic><topic>Amino Acid Isomerases - isolation & purification</topic><topic>Amino Acid Isomerases - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern</topic><topic>Carrier Proteins - genetics</topic><topic>Carrier Proteins - isolation & purification</topic><topic>Carrier Proteins - metabolism</topic><topic>cDNA</topic><topic>Cloning, Molecular</topic><topic>cyclophilin</topic><topic>Cytosol - enzymology</topic><topic>DNA - genetics</topic><topic>DNA - isolation & purification</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Library</topic><topic>genes</topic><topic>Genes. Genome</topic><topic>Kinetics</topic><topic>Lycopersicon esculentum</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>nucleotide sequence</topic><topic>Oligonucleotide Probes</topic><topic>Peptidylprolyl Isomerase</topic><topic>Plants - enzymology</topic><topic>Plants - genetics</topic><topic>predictions</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>RNA - genetics</topic><topic>RNA - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GASSER, C. S</creatorcontrib><creatorcontrib>GUNNING, D. A</creatorcontrib><creatorcontrib>BUDELIER, K. A</creatorcontrib><creatorcontrib>BROWN, S. M</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</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>GASSER, C. S</au><au>GUNNING, D. A</au><au>BUDELIER, K. A</au><au>BROWN, S. M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1990-12-01</date><risdate>1990</risdate><volume>87</volume><issue>24</issue><spage>9519</spage><epage>9523</epage><pages>9519-9523</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>cDNA clones encoding proteins of approximately 18 kDa in which 83% of the amino acids are conserved relative to the published sequences of mammalian cyclophilin/rotamase (CyP) have been isolated from tomato, maize, and Brassica napus. In correspondence with the mammalian genes, but in contrast with the Neurospora gene and one yeast CyP gene, the plant CyP genes encode only mature proteins lacking transit peptides. RNA blot analyses demonstrate that CyP genes are expressed in all plant organs tested. Southern blots of genomic DNA indicate that there are small families (two to eight members) of CyP-related genes in maize and B. napus. A vector was constructed for expression of the tomato cDNA in E. coli. SDS/polyacrylamide gels show that extracts of appropriately induced cells harboring this vector contain nearly 40% of the protein as a single approximately 18-kDa band. While the majority of this protein is sequestered in insoluble inclusion bodies, the soluble extracts have higher levels of peptidyl-prolyl cis-trans isomerase (rotamase) activity than extracts of wild-type cells. This additional activity is sensitive to inhibition by the cyclic undecapeptide cyclosporin A.</abstract><cop>Washington, DC</cop><pub>National Acad Sciences</pub><pmid>1702215</pmid><doi>10.1073/pnas.87.24.9519</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1990-12, Vol.87 (24), p.9519-9523 |
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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 | Amino Acid Isomerases - genetics Amino Acid Isomerases - isolation & purification Amino Acid Isomerases - metabolism Amino Acid Sequence Base Sequence Biological and medical sciences Blotting, Southern Carrier Proteins - genetics Carrier Proteins - isolation & purification Carrier Proteins - metabolism cDNA Cloning, Molecular cyclophilin Cytosol - enzymology DNA - genetics DNA - isolation & purification Electrophoresis, Polyacrylamide Gel Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Library genes Genes. Genome Kinetics Lycopersicon esculentum Molecular and cellular biology Molecular genetics Molecular Sequence Data Molecular Weight nucleotide sequence Oligonucleotide Probes Peptidylprolyl Isomerase Plants - enzymology Plants - genetics predictions Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism RNA - genetics RNA - isolation & purification |
| title | Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli |
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