Ultrasensitive detection of protein translocated through toxin pores in droplet-interface bilayers
Many bacterial toxins form proteinaceous pores that facilitate the translocation of soluble effector proteins across cellular membranes. With anthrax toxin this process may be monitored in real time by electrophysiology, where fluctuations in ionic current through these pores inserted in model membr...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2011-10, Vol.108 (40), p.16577-16581 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Fischer, Audrey Holden, Matthew A Pentelute, Brad L Collier, R. John |
| description | Many bacterial toxins form proteinaceous pores that facilitate the translocation of soluble effector proteins across cellular membranes. With anthrax toxin this process may be monitored in real time by electrophysiology, where fluctuations in ionic current through these pores inserted in model membranes are used to infer the translocation of individual protein molecules. However, detecting the minute quantities of translocated proteins has been a challenge. Here, we describe use of the droplet-interface bilayer system to follow the movement of proteins across a model membrane separating two submicroliter aqueous droplets. We report the capture and subsequent direct detection of as few as 100 protein molecules that have translocated through anthrax toxin pores. The droplet-interface bilayer system offers new avenues of approach to the study of protein translocation. |
| doi_str_mv | 10.1073/pnas.1113074108 |
| format | Article |
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The droplet-interface bilayer system offers new avenues of approach to the study of protein translocation.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1113074108</identifier><identifier>PMID: 21949363</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Anthrax ; Antigens ; Antigens, Bacterial - chemistry ; Antigens, Bacterial - metabolism ; bacterial toxins ; Bacterial Toxins - chemistry ; Bacterial Toxins - metabolism ; Biological Sciences ; Carrier Proteins - metabolism ; droplets ; Electric potential ; Electrodes ; Electrophysiology - methods ; Freight ; Lipid Bilayers - metabolism ; Membrane potential ; Membranes ; Molecules ; Pore Forming Cytotoxic Proteins - chemistry ; Pore Forming Cytotoxic Proteins - metabolism ; Protein transport ; Protein Transport - physiology ; Proteins ; Toxins</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2011-10, Vol.108 (40), p.16577-16581</ispartof><rights>copyright © 1993-2008 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Oct 4, 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c589t-ddf8612122bb09c0bb3d1d3d0eb08b12b271e72a4959ce55163b8929478c253f3</citedby><cites>FETCH-LOGICAL-c589t-ddf8612122bb09c0bb3d1d3d0eb08b12b271e72a4959ce55163b8929478c253f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/108/40.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/41321743$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/41321743$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27924,27925,53791,53793,58017,58250</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21949363$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fischer, Audrey</creatorcontrib><creatorcontrib>Holden, Matthew A</creatorcontrib><creatorcontrib>Pentelute, Brad L</creatorcontrib><creatorcontrib>Collier, R. John</creatorcontrib><title>Ultrasensitive detection of protein translocated through toxin pores in droplet-interface bilayers</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Many bacterial toxins form proteinaceous pores that facilitate the translocation of soluble effector proteins across cellular membranes. With anthrax toxin this process may be monitored in real time by electrophysiology, where fluctuations in ionic current through these pores inserted in model membranes are used to infer the translocation of individual protein molecules. However, detecting the minute quantities of translocated proteins has been a challenge. Here, we describe use of the droplet-interface bilayer system to follow the movement of proteins across a model membrane separating two submicroliter aqueous droplets. We report the capture and subsequent direct detection of as few as 100 protein molecules that have translocated through anthrax toxin pores. The droplet-interface bilayer system offers new avenues of approach to the study of protein translocation.</description><subject>Anthrax</subject><subject>Antigens</subject><subject>Antigens, Bacterial - chemistry</subject><subject>Antigens, Bacterial - metabolism</subject><subject>bacterial toxins</subject><subject>Bacterial Toxins - chemistry</subject><subject>Bacterial Toxins - metabolism</subject><subject>Biological Sciences</subject><subject>Carrier Proteins - metabolism</subject><subject>droplets</subject><subject>Electric potential</subject><subject>Electrodes</subject><subject>Electrophysiology - methods</subject><subject>Freight</subject><subject>Lipid Bilayers - metabolism</subject><subject>Membrane potential</subject><subject>Membranes</subject><subject>Molecules</subject><subject>Pore Forming Cytotoxic Proteins - chemistry</subject><subject>Pore Forming Cytotoxic Proteins - metabolism</subject><subject>Protein transport</subject><subject>Protein Transport - physiology</subject><subject>Proteins</subject><subject>Toxins</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kk2P0zAQhi0EYkvhzAmIuMAlux7bcewLElrxJa3EAXq27GTSukrjYLsr9t_jqqULHDjZ0jzzaDyvCXkO9BJoy6_myaZLAOC0FUDVA7IAqqGWQtOHZEEpa2slmLggT1LaUkp1o-hjcsFAC80lXxC3GnO0Cafks7_FqseMXfZhqsJQzTFk9FNViCmNobMZ-ypvYtivN1UOP0tpDhFTVS59DPOIufZTxjjYDivnR3uHMT0ljwY7Jnx2Opdk9fHD9-vP9c3XT1-u39_UXaN0rvt-UBIYMOYc1R11jvfQ856io8oBc6wFbJkVutEdNg1I7pRmWrSqYw0f-JK8O3rnvdth3-FU5h7NHP3OxjsTrDd_Vya_MetwazgoTaUugjcnQQw_9piy2fnU4TjaCcM-GaVlw4Qu216St_8lgTdSMwmtLOjrf9Bt2MepLOLgk7rEQAt0dYS6GFKKOJynBmoOQZtD0OY-6NLx8s_HnvnfyRagOgGHznudMqIoZdO2BXlxRLYph3hmBHAGrTgoXh3rgw3GrqNPZvWNURDlI0ld9s9_AQ1-w1U</recordid><startdate>20111004</startdate><enddate>20111004</enddate><creator>Fischer, Audrey</creator><creator>Holden, Matthew A</creator><creator>Pentelute, Brad L</creator><creator>Collier, R. John</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</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>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>7X8</scope><scope>5PM</scope></search><sort><creationdate>20111004</creationdate><title>Ultrasensitive detection of protein translocated through toxin pores in droplet-interface bilayers</title><author>Fischer, Audrey ; Holden, Matthew A ; Pentelute, Brad L ; Collier, R. 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| subjects | Anthrax Antigens Antigens, Bacterial - chemistry Antigens, Bacterial - metabolism bacterial toxins Bacterial Toxins - chemistry Bacterial Toxins - metabolism Biological Sciences Carrier Proteins - metabolism droplets Electric potential Electrodes Electrophysiology - methods Freight Lipid Bilayers - metabolism Membrane potential Membranes Molecules Pore Forming Cytotoxic Proteins - chemistry Pore Forming Cytotoxic Proteins - metabolism Protein transport Protein Transport - physiology Proteins Toxins |
| title | Ultrasensitive detection of protein translocated through toxin pores in droplet-interface bilayers |
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