Outer‐Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate
E. coli and Salmonella are two of the most common bacterial pathogens involved in foodborne and waterborne related deaths. Hence, it is critical to develop rapid and sensitive detection strategies for near‐outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL−1...
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| Veröffentlicht in: | Angewandte Chemie International Edition 2020-10, Vol.59 (41), p.18068-18077 |
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| creator | Sinsinbar, Gaurav Gudlur, Sushanth Wood, Sarah E. Ammanath, Gopal Yildiz, Hakan U. Alagappan, Palaniappan Mrksich, Milan Liedberg, Bo |
| description | E. coli and Salmonella are two of the most common bacterial pathogens involved in foodborne and waterborne related deaths. Hence, it is critical to develop rapid and sensitive detection strategies for near‐outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL−1 of E. coli in water within 6 hours by targeting the bacteria's surface protease activity. The assay relies on polythiophene acetic acid (PTAA) as an optical reporter and a short unlabeled peptide (LL37FRRV) previously optimized as a substrate for OmpT, an outer‐membrane protease on E. coli. LL37FRRV interacts with PTAA to enhance its fluorescence while also inducing the formation of a helical PTAA‐LL37FRRV construct, as confirmed by circular dichroism. However, in the presence of E. coli LL37FRRV is cleaved and can no longer affect the conformations and optical properties of PTAA. This ability to distinguish between an intact and cleaved peptide was investigated in detail using LL37FRRV sequence variants.
The structural and optical changes resulting from the interaction between a polythiophene (PTAA) and a short unlabeled peptide were exploited to develop a sensitive bacterial detection method. The method itself targets the bacteria's surface protease activity and is sensitive enough to detect 1 CFU mL−1 of E. coli in water within 6 hours. |
| doi_str_mv | 10.1002/anie.202008444 |
| format | Article |
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The structural and optical changes resulting from the interaction between a polythiophene (PTAA) and a short unlabeled peptide were exploited to develop a sensitive bacterial detection method. The method itself targets the bacteria's surface protease activity and is sensitive enough to detect 1 CFU mL−1 of E. coli in water within 6 hours.</description><edition>International ed. in English</edition><identifier>ISSN: 1433-7851</identifier><identifier>EISSN: 1521-3773</identifier><identifier>DOI: 10.1002/anie.202008444</identifier><identifier>PMID: 32618102</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>Acetic acid ; Amino Acid Sequence ; Anions ; Bacterial Outer Membrane Proteins - chemistry ; Bacterial Outer Membrane Proteins - metabolism ; Circular dichroism ; Colony Count, Microbial ; Dichroism ; E coli ; enzymes ; Escherichia coli ; Escherichia coli - isolation & purification ; Escherichia coli - metabolism ; Escherichia coli Proteins - chemistry ; Escherichia coli Proteins - metabolism ; Fluorescence ; Foodborne diseases ; membrane proteins ; Membranes ; Optical properties ; pathogens ; Peptide Hydrolases - chemistry ; Peptide Hydrolases - metabolism ; Peptides ; Peptides - metabolism ; Polymers - metabolism ; Polythiophene ; Protease ; Proteinase ; Salmonella ; Spectrometry, Fluorescence ; Substrate Specificity ; Substrates ; Thiophenes - metabolism ; Water Microbiology ; Waterborne diseases</subject><ispartof>Angewandte Chemie International Edition, 2020-10, Vol.59 (41), p.18068-18077</ispartof><rights>2020 Wiley‐VCH GmbH</rights><rights>2020 Wiley-VCH GmbH.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3654-8a381fbf00169139bf81d4b2a8dda95366428fe9984a2134d691827d0d4ed8493</citedby><cites>FETCH-LOGICAL-c3654-8a381fbf00169139bf81d4b2a8dda95366428fe9984a2134d691827d0d4ed8493</cites><orcidid>0000-0003-2883-6953</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fanie.202008444$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fanie.202008444$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32618102$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sinsinbar, Gaurav</creatorcontrib><creatorcontrib>Gudlur, Sushanth</creatorcontrib><creatorcontrib>Wood, Sarah E.</creatorcontrib><creatorcontrib>Ammanath, Gopal</creatorcontrib><creatorcontrib>Yildiz, Hakan U.</creatorcontrib><creatorcontrib>Alagappan, Palaniappan</creatorcontrib><creatorcontrib>Mrksich, Milan</creatorcontrib><creatorcontrib>Liedberg, Bo</creatorcontrib><title>Outer‐Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate</title><title>Angewandte Chemie International Edition</title><addtitle>Angew Chem Int Ed Engl</addtitle><description>E. coli and Salmonella are two of the most common bacterial pathogens involved in foodborne and waterborne related deaths. Hence, it is critical to develop rapid and sensitive detection strategies for near‐outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL−1 of E. coli in water within 6 hours by targeting the bacteria's surface protease activity. The assay relies on polythiophene acetic acid (PTAA) as an optical reporter and a short unlabeled peptide (LL37FRRV) previously optimized as a substrate for OmpT, an outer‐membrane protease on E. coli. LL37FRRV interacts with PTAA to enhance its fluorescence while also inducing the formation of a helical PTAA‐LL37FRRV construct, as confirmed by circular dichroism. However, in the presence of E. coli LL37FRRV is cleaved and can no longer affect the conformations and optical properties of PTAA. This ability to distinguish between an intact and cleaved peptide was investigated in detail using LL37FRRV sequence variants.
The structural and optical changes resulting from the interaction between a polythiophene (PTAA) and a short unlabeled peptide were exploited to develop a sensitive bacterial detection method. The method itself targets the bacteria's surface protease activity and is sensitive enough to detect 1 CFU mL−1 of E. coli in water within 6 hours.</description><subject>Acetic acid</subject><subject>Amino Acid Sequence</subject><subject>Anions</subject><subject>Bacterial Outer Membrane Proteins - chemistry</subject><subject>Bacterial Outer Membrane Proteins - metabolism</subject><subject>Circular dichroism</subject><subject>Colony Count, Microbial</subject><subject>Dichroism</subject><subject>E coli</subject><subject>enzymes</subject><subject>Escherichia coli</subject><subject>Escherichia coli - isolation & purification</subject><subject>Escherichia coli - metabolism</subject><subject>Escherichia coli Proteins - chemistry</subject><subject>Escherichia coli Proteins - metabolism</subject><subject>Fluorescence</subject><subject>Foodborne diseases</subject><subject>membrane proteins</subject><subject>Membranes</subject><subject>Optical properties</subject><subject>pathogens</subject><subject>Peptide Hydrolases - chemistry</subject><subject>Peptide Hydrolases - metabolism</subject><subject>Peptides</subject><subject>Peptides - metabolism</subject><subject>Polymers - metabolism</subject><subject>Polythiophene</subject><subject>Protease</subject><subject>Proteinase</subject><subject>Salmonella</subject><subject>Spectrometry, Fluorescence</subject><subject>Substrate Specificity</subject><subject>Substrates</subject><subject>Thiophenes - metabolism</subject><subject>Water Microbiology</subject><subject>Waterborne diseases</subject><issn>1433-7851</issn><issn>1521-3773</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0U1P2zAYB3Br2jRetivHydIu7JDityTOsVQdIDFaqeUcOfETapTYwU5U9cZH4DPySTAqMGmXnfwcfs9ftv8InVAyoYSwM2UNTBhhhEghxCd0SFNGE57n_HOcBedJLlN6gI5CuI9eSpJ9RQecZVRSwg7Rw2IcwD8_Pv2BrvLKAl56N4AKgE8XXb_-hc_jrPF8gmvXGrwCG4y9w1szbPDUGmdNjZeu3Q0b4_oNxABlNb61raqgjYtL6AejAa_GKgxeDfANfWlUG-D723mMbn_P17PL5HpxcTWbXic1z1KRSMUlbaqGEJoVlBdVI6kWFVNSa1WkPMsEkw0UhRSKUS50VJLlmmgBWoqCH6PTfW7v3cMIYSg7E2po2_hIN4aSCUYoT7kkkf78h9670dt4u6hEllOZkTyqyV7V3oXgoSl7bzrldyUl5WsZ5WsZ5UcZceHHW-xYdaA_-PvvR1Dswda0sPtPXDm9uZr_DX8BhnqVRw</recordid><startdate>20201005</startdate><enddate>20201005</enddate><creator>Sinsinbar, Gaurav</creator><creator>Gudlur, Sushanth</creator><creator>Wood, Sarah E.</creator><creator>Ammanath, Gopal</creator><creator>Yildiz, Hakan U.</creator><creator>Alagappan, Palaniappan</creator><creator>Mrksich, Milan</creator><creator>Liedberg, Bo</creator><general>Wiley Subscription Services, Inc</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>7TM</scope><scope>K9.</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-2883-6953</orcidid></search><sort><creationdate>20201005</creationdate><title>Outer‐Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate</title><author>Sinsinbar, Gaurav ; 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Hence, it is critical to develop rapid and sensitive detection strategies for near‐outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL−1 of E. coli in water within 6 hours by targeting the bacteria's surface protease activity. The assay relies on polythiophene acetic acid (PTAA) as an optical reporter and a short unlabeled peptide (LL37FRRV) previously optimized as a substrate for OmpT, an outer‐membrane protease on E. coli. LL37FRRV interacts with PTAA to enhance its fluorescence while also inducing the formation of a helical PTAA‐LL37FRRV construct, as confirmed by circular dichroism. However, in the presence of E. coli LL37FRRV is cleaved and can no longer affect the conformations and optical properties of PTAA. This ability to distinguish between an intact and cleaved peptide was investigated in detail using LL37FRRV sequence variants.
The structural and optical changes resulting from the interaction between a polythiophene (PTAA) and a short unlabeled peptide were exploited to develop a sensitive bacterial detection method. The method itself targets the bacteria's surface protease activity and is sensitive enough to detect 1 CFU mL−1 of E. coli in water within 6 hours.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>32618102</pmid><doi>10.1002/anie.202008444</doi><tpages>10</tpages><edition>International ed. in English</edition><orcidid>https://orcid.org/0000-0003-2883-6953</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Acetic acid Amino Acid Sequence Anions Bacterial Outer Membrane Proteins - chemistry Bacterial Outer Membrane Proteins - metabolism Circular dichroism Colony Count, Microbial Dichroism E coli enzymes Escherichia coli Escherichia coli - isolation & purification Escherichia coli - metabolism Escherichia coli Proteins - chemistry Escherichia coli Proteins - metabolism Fluorescence Foodborne diseases membrane proteins Membranes Optical properties pathogens Peptide Hydrolases - chemistry Peptide Hydrolases - metabolism Peptides Peptides - metabolism Polymers - metabolism Polythiophene Protease Proteinase Salmonella Spectrometry, Fluorescence Substrate Specificity Substrates Thiophenes - metabolism Water Microbiology Waterborne diseases |
| title | Outer‐Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate |
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