Retrograde Transport of Mutant Ricin to the Endoplasmic Reticulum with Subsequent Translocation to Cytosol
Translocation of ricin A chain to the cytosol has been proposed to take place from the endoplasmic reticulum (ER), but attempts to visualize ricin in this organelle have failed. Here we modified ricin A chain to contain a tyrosine sulfation site alone or in combination with N-glycosylation sites. Wh...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1997-04, Vol.94 (8), p.3783-3788 |
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| creator | Rapak, Andrzej Falnes, Pal O. Olsnes, Sjur |
| description | Translocation of ricin A chain to the cytosol has been proposed to take place from the endoplasmic reticulum (ER), but attempts to visualize ricin in this organelle have failed. Here we modified ricin A chain to contain a tyrosine sulfation site alone or in combination with N-glycosylation sites. When reconstituted with ricin B chain and incubated with cells in the presence of Na2
35SO4, the modified A chains were labeled. The labeling was prevented by brefeldin A and ilimaquinone, and it appears to take place in the Golgi apparatus. This method allows selective labeling of ricin molecules that have already been transported retrograde to this organelle. A chain containing C-terminal N-glycosylation sites became core glycosylated, indicating retrograde transport to the ER. In part of the toxin molecules, the A chain was released from the B chain and translocated to the cytosol. The finding that glycosylated A chain was present in the cytosol indicates that translocation takes place after transport of the toxin to the ER. |
| doi_str_mv | 10.1073/pnas.94.8.3783 |
| format | Article |
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35SO4, the modified A chains were labeled. The labeling was prevented by brefeldin A and ilimaquinone, and it appears to take place in the Golgi apparatus. This method allows selective labeling of ricin molecules that have already been transported retrograde to this organelle. A chain containing C-terminal N-glycosylation sites became core glycosylated, indicating retrograde transport to the ER. In part of the toxin molecules, the A chain was released from the B chain and translocated to the cytosol. The finding that glycosylated A chain was present in the cytosol indicates that translocation takes place after transport of the toxin to the ER.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.94.8.3783</identifier><identifier>PMID: 9108055</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Animals ; Antibodies ; Base Sequence ; Biological Sciences ; Biological Transport ; Cell membranes ; Cellular biology ; Cercopithecus aethiops ; Cytosol ; Cytosol - metabolism ; Endoplasmic Reticulum - metabolism ; Enzymes ; Golgi apparatus ; Golgi Apparatus - metabolism ; Molecular Sequence Data ; Molecules ; Mutation ; Organelles ; Plasmids ; Proteins ; Ricin - genetics ; Ricin - metabolism ; Sulfation ; Toxicity ; Toxins ; Vero Cells</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1997-04, Vol.94 (8), p.3783-3788</ispartof><rights>Copyright 1997 National Academy of Sciences</rights><rights>Copyright National Academy of Sciences Apr 15, 1997</rights><rights>Copyright © 1997, The National Academy of Sciences of the USA 1997</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c513t-593712e407786bd70bca93ca9f787541b286b4b0929c567bbdb29e1307520ebf3</citedby><cites>FETCH-LOGICAL-c513t-593712e407786bd70bca93ca9f787541b286b4b0929c567bbdb29e1307520ebf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/94/8.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/41907$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/41907$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,725,778,782,801,883,27911,27912,53778,53780,58004,58237</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9108055$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rapak, Andrzej</creatorcontrib><creatorcontrib>Falnes, Pal O.</creatorcontrib><creatorcontrib>Olsnes, Sjur</creatorcontrib><title>Retrograde Transport of Mutant Ricin to the Endoplasmic Reticulum with Subsequent Translocation to Cytosol</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Translocation of ricin A chain to the cytosol has been proposed to take place from the endoplasmic reticulum (ER), but attempts to visualize ricin in this organelle have failed. Here we modified ricin A chain to contain a tyrosine sulfation site alone or in combination with N-glycosylation sites. When reconstituted with ricin B chain and incubated with cells in the presence of Na2
35SO4, the modified A chains were labeled. The labeling was prevented by brefeldin A and ilimaquinone, and it appears to take place in the Golgi apparatus. This method allows selective labeling of ricin molecules that have already been transported retrograde to this organelle. A chain containing C-terminal N-glycosylation sites became core glycosylated, indicating retrograde transport to the ER. In part of the toxin molecules, the A chain was released from the B chain and translocated to the cytosol. The finding that glycosylated A chain was present in the cytosol indicates that translocation takes place after transport of the toxin to the ER.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Base Sequence</subject><subject>Biological Sciences</subject><subject>Biological Transport</subject><subject>Cell membranes</subject><subject>Cellular biology</subject><subject>Cercopithecus aethiops</subject><subject>Cytosol</subject><subject>Cytosol - metabolism</subject><subject>Endoplasmic Reticulum - metabolism</subject><subject>Enzymes</subject><subject>Golgi apparatus</subject><subject>Golgi Apparatus - metabolism</subject><subject>Molecular Sequence Data</subject><subject>Molecules</subject><subject>Mutation</subject><subject>Organelles</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Ricin - genetics</subject><subject>Ricin - metabolism</subject><subject>Sulfation</subject><subject>Toxicity</subject><subject>Toxins</subject><subject>Vero Cells</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc9rFDEYhoModa1ePQhC8NDbjF_mVxLwIktthYpQ6zkkmUw3S2YyJhm1_73Z7rKsHjyEQN7n-XjDh9BrAiUBWr-fJxlL3pSsrCmrn6AVAU6KruHwFK0AKlqwpmqeoxcxbgGAtwzO0BknwKBtV2h7a1Lw90H2Bt8FOcXZh4T9gL8sSU4J31ptJ5w8ThuDL6fez07G0WqcPasXt4z4l00b_G1R0fxYTFYexzivZbL-UV0_JB-9e4meDdJF8-pwn6Pvny7v1tfFzderz-uPN4VuSZ2KlteUVKYBSlmnegpKS17nM1BG24aoKj83CnjFddtRpXpVcUNqoG0FRg31OfqwnzsvajS9zp2CdGIOdpThQXhpxd_JZDfi3v8UFbSEZf3ioAefPxSTGG3Uxjk5Gb9EQToAwijP4Lt_wK1fwpS_liflPjXrIEPlHtLBxxjMcOxBQOwWKHYLFLwRTOwWmIW3p-2P-GFjJ_nOO6Yn_sX_cjEsziXzO2XwzR7cxuTDkWwIz63-AO-xuaA</recordid><startdate>19970415</startdate><enddate>19970415</enddate><creator>Rapak, Andrzej</creator><creator>Falnes, Pal O.</creator><creator>Olsnes, Sjur</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><general>The National Academy of Sciences of the USA</general><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>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7U7</scope><scope>5PM</scope></search><sort><creationdate>19970415</creationdate><title>Retrograde Transport of Mutant Ricin to the Endoplasmic Reticulum with Subsequent Translocation to Cytosol</title><author>Rapak, Andrzej ; Falnes, Pal O. ; Olsnes, Sjur</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c513t-593712e407786bd70bca93ca9f787541b286b4b0929c567bbdb29e1307520ebf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Base Sequence</topic><topic>Biological Sciences</topic><topic>Biological Transport</topic><topic>Cell membranes</topic><topic>Cellular biology</topic><topic>Cercopithecus aethiops</topic><topic>Cytosol</topic><topic>Cytosol - metabolism</topic><topic>Endoplasmic Reticulum - metabolism</topic><topic>Enzymes</topic><topic>Golgi apparatus</topic><topic>Golgi Apparatus - metabolism</topic><topic>Molecular Sequence Data</topic><topic>Molecules</topic><topic>Mutation</topic><topic>Organelles</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Ricin - genetics</topic><topic>Ricin - metabolism</topic><topic>Sulfation</topic><topic>Toxicity</topic><topic>Toxins</topic><topic>Vero Cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rapak, Andrzej</creatorcontrib><creatorcontrib>Falnes, Pal O.</creatorcontrib><creatorcontrib>Olsnes, Sjur</creatorcontrib><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>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Toxicology Abstracts</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>Rapak, Andrzej</au><au>Falnes, Pal O.</au><au>Olsnes, Sjur</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Retrograde Transport of Mutant Ricin to the Endoplasmic Reticulum with Subsequent Translocation to Cytosol</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1997-04-15</date><risdate>1997</risdate><volume>94</volume><issue>8</issue><spage>3783</spage><epage>3788</epage><pages>3783-3788</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Translocation of ricin A chain to the cytosol has been proposed to take place from the endoplasmic reticulum (ER), but attempts to visualize ricin in this organelle have failed. Here we modified ricin A chain to contain a tyrosine sulfation site alone or in combination with N-glycosylation sites. When reconstituted with ricin B chain and incubated with cells in the presence of Na2
35SO4, the modified A chains were labeled. The labeling was prevented by brefeldin A and ilimaquinone, and it appears to take place in the Golgi apparatus. This method allows selective labeling of ricin molecules that have already been transported retrograde to this organelle. A chain containing C-terminal N-glycosylation sites became core glycosylated, indicating retrograde transport to the ER. In part of the toxin molecules, the A chain was released from the B chain and translocated to the cytosol. The finding that glycosylated A chain was present in the cytosol indicates that translocation takes place after transport of the toxin to the ER.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>9108055</pmid><doi>10.1073/pnas.94.8.3783</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1997-04, Vol.94 (8), p.3783-3788 |
| issn | 0027-8424 1091-6490 |
| language | eng |
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| subjects | Animals Antibodies Base Sequence Biological Sciences Biological Transport Cell membranes Cellular biology Cercopithecus aethiops Cytosol Cytosol - metabolism Endoplasmic Reticulum - metabolism Enzymes Golgi apparatus Golgi Apparatus - metabolism Molecular Sequence Data Molecules Mutation Organelles Plasmids Proteins Ricin - genetics Ricin - metabolism Sulfation Toxicity Toxins Vero Cells |
| title | Retrograde Transport of Mutant Ricin to the Endoplasmic Reticulum with Subsequent Translocation to Cytosol |
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