A modified fluorescent sensor for reporting glucose concentration in the airway lumen

We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP la...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2021-07, Vol.16 (7), p.e0254248
Main Authors: Bearham, Jade, Krutrök, Nina, Lindberg, Botilda, Woodall, Maximillian, Astrand, Annika, Taylor, John D, Biggart, Matthew, Vasiljevs, Stanislavs, Tarran, Robert, Baines, Deborah L
Format: Article
Language:English
Subjects:
Analysis
Animals
Biology and Life Sciences
Biosensing Techniques - methods
Bronchus
Calcium-Binding Proteins
Chemical compounds
Dextrose
Diabetes
Diabetes mellitus
E coli
Engineering and Technology
Epithelial cells
Epithelium
Escherichia coli
Escherichia coli - metabolism
Ethylenediamine
Fluorescence
Fluorescent Dyes - chemistry
Fluorophores
Galactose
Genomics
Glucose
Glucose - metabolism
Hemodialysis
Humans
Immunology
Inhalation
Lungs
Measurement
Medicine and Health Sciences
Mice
Mice, Inbred C57BL
Molecular structure
Monosaccharide Transport Proteins - metabolism
Mutagenesis
Mutation
Periplasmic Binding Proteins - metabolism
Physical Sciences
Plasmids
Protein expression
Proteins
R&D
Research & development
Research and Analysis Methods
Respiration
Respiratory Mucosa - metabolism
Respiratory tract
Sensors
Staphylococcus infections
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3
cites cdi_FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3
container_end_page
container_issue 7
container_start_page e0254248
container_title PloS one
container_volume 16
creator Bearham, Jade
Krutrök, Nina
Lindberg, Botilda
Woodall, Maximillian
Astrand, Annika
Taylor, John D
Biggart, Matthew
Vasiljevs, Stanislavs
Tarran, Robert
Baines, Deborah L
description We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N'-Dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations
doi_str_mv 10.1371/journal.pone.0254248
format article
fullrecord <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_2549936790</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A667980252</galeid><doaj_id>oai_doaj_org_article_6fa398a5f63244c588c16a9063879c43</doaj_id><sourcerecordid>A667980252</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3</originalsourceid><addsrcrecordid>eNqNkluL1DAYhoso7kH_gWhBWPRixhzaNLkRhsXDwMKCut6GTA6dDGkyJq26_97U6S5T2QspoSV5vjf93u8tihcQLCFu4LtdGKIXbrkPXi8BqitU0UfFKWQYLQgC-PHR90lxltIOgBpTQp4WJzjDCDF0Wtysyi4oa6xWpXFDiDpJ7fsyaZ9CLE1eUe9D7K1vy9YNMiRdyuBHKIreBl9aX_ZbXQobf4nb0g2d9s-KJ0a4pJ9P7_Pi5uOHb5efF1fXn9aXq6uFJAz1CwTJRmxoxTCpKaspRAAoBRqqpUIYgAZKA5FEoqkBpUqajcFGKAY3UCqtFT4vXh109y4kPjmSeDaDZc2GgUysD4QKYsf30XYi3vIgLP-7EWLLRW5OOs2JEZhRURuCUVXJmlIJiWCAYNowWeGs9X66bdh0Wh0scDPR-Ym3W96Gn5yiBsCmyQJvJoEYfgw69byz2W7nhNdhGP-7zoNklLCMvv4Hfbi7iWpFbsB6E_K9chTlK5IJmuVQppYPUPlRurN5ltrYvD8reDsryEyvf_etGFLi669f_p-9_j5nL47YrRau36bghjFGaQ5WB1DGkFLU5t5kCPiY_js3-Jh-PqU_l708HtB90V3c8R92Uf9F</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2549936790</pqid></control><display><type>article</type><title>A modified fluorescent sensor for reporting glucose concentration in the airway lumen</title><source>Publicly Available Content Database</source><source>PubMed Central</source><creator>Bearham, Jade ; Krutrök, Nina ; Lindberg, Botilda ; Woodall, Maximillian ; Astrand, Annika ; Taylor, John D ; Biggart, Matthew ; Vasiljevs, Stanislavs ; Tarran, Robert ; Baines, Deborah L</creator><contributor>Koval, Michael</contributor><creatorcontrib>Bearham, Jade ; Krutrök, Nina ; Lindberg, Botilda ; Woodall, Maximillian ; Astrand, Annika ; Taylor, John D ; Biggart, Matthew ; Vasiljevs, Stanislavs ; Tarran, Robert ; Baines, Deborah L ; Koval, Michael</creatorcontrib><description>We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N'-Dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations &lt;50μM. Using nasal inhalation of the sensor and spectral unmixing to generate images, E149C/A213R GBP-IANBD fluorescence was detected in luminal regions of cryosections of the murine distal lung that was greater in db/db than wildtype mice. In conclusion, this sensor provides a useful tool for further development to measure luminal glucose concentration in models of lung/airway to explore how this may change in disease.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0254248</identifier><identifier>PMID: 34242292</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Animals ; Biology and Life Sciences ; Biosensing Techniques - methods ; Bronchus ; Calcium-Binding Proteins ; Chemical compounds ; Dextrose ; Diabetes ; Diabetes mellitus ; E coli ; Engineering and Technology ; Epithelial cells ; Epithelium ; Escherichia coli ; Escherichia coli - metabolism ; Ethylenediamine ; Fluorescence ; Fluorescent Dyes - chemistry ; Fluorophores ; Galactose ; Genomics ; Glucose ; Glucose - metabolism ; Hemodialysis ; Humans ; Immunology ; Inhalation ; Lungs ; Measurement ; Medicine and Health Sciences ; Mice ; Mice, Inbred C57BL ; Molecular structure ; Monosaccharide Transport Proteins - metabolism ; Mutagenesis ; Mutation ; Periplasmic Binding Proteins - metabolism ; Physical Sciences ; Plasmids ; Protein expression ; Proteins ; R&amp;D ; Research &amp; development ; Research and Analysis Methods ; Respiration ; Respiratory Mucosa - metabolism ; Respiratory tract ; Sensors ; Staphylococcus infections</subject><ispartof>PloS one, 2021-07, Vol.16 (7), p.e0254248</ispartof><rights>COPYRIGHT 2021 Public Library of Science</rights><rights>2021 Bearham et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 Bearham et al 2021 Bearham et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3</citedby><cites>FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3</cites><orcidid>0000-0001-7229-0244</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2549936790/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2549936790?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34242292$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Koval, Michael</contributor><creatorcontrib>Bearham, Jade</creatorcontrib><creatorcontrib>Krutrök, Nina</creatorcontrib><creatorcontrib>Lindberg, Botilda</creatorcontrib><creatorcontrib>Woodall, Maximillian</creatorcontrib><creatorcontrib>Astrand, Annika</creatorcontrib><creatorcontrib>Taylor, John D</creatorcontrib><creatorcontrib>Biggart, Matthew</creatorcontrib><creatorcontrib>Vasiljevs, Stanislavs</creatorcontrib><creatorcontrib>Tarran, Robert</creatorcontrib><creatorcontrib>Baines, Deborah L</creatorcontrib><title>A modified fluorescent sensor for reporting glucose concentration in the airway lumen</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N'-Dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations &lt;50μM. Using nasal inhalation of the sensor and spectral unmixing to generate images, E149C/A213R GBP-IANBD fluorescence was detected in luminal regions of cryosections of the murine distal lung that was greater in db/db than wildtype mice. In conclusion, this sensor provides a useful tool for further development to measure luminal glucose concentration in models of lung/airway to explore how this may change in disease.</description><subject>Analysis</subject><subject>Animals</subject><subject>Biology and Life Sciences</subject><subject>Biosensing Techniques - methods</subject><subject>Bronchus</subject><subject>Calcium-Binding Proteins</subject><subject>Chemical compounds</subject><subject>Dextrose</subject><subject>Diabetes</subject><subject>Diabetes mellitus</subject><subject>E coli</subject><subject>Engineering and Technology</subject><subject>Epithelial cells</subject><subject>Epithelium</subject><subject>Escherichia coli</subject><subject>Escherichia coli - metabolism</subject><subject>Ethylenediamine</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes - chemistry</subject><subject>Fluorophores</subject><subject>Galactose</subject><subject>Genomics</subject><subject>Glucose</subject><subject>Glucose - metabolism</subject><subject>Hemodialysis</subject><subject>Humans</subject><subject>Immunology</subject><subject>Inhalation</subject><subject>Lungs</subject><subject>Measurement</subject><subject>Medicine and Health Sciences</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Molecular structure</subject><subject>Monosaccharide Transport Proteins - metabolism</subject><subject>Mutagenesis</subject><subject>Mutation</subject><subject>Periplasmic Binding Proteins - metabolism</subject><subject>Physical Sciences</subject><subject>Plasmids</subject><subject>Protein expression</subject><subject>Proteins</subject><subject>R&amp;D</subject><subject>Research &amp; development</subject><subject>Research and Analysis Methods</subject><subject>Respiration</subject><subject>Respiratory Mucosa - metabolism</subject><subject>Respiratory tract</subject><subject>Sensors</subject><subject>Staphylococcus infections</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNqNkluL1DAYhoso7kH_gWhBWPRixhzaNLkRhsXDwMKCut6GTA6dDGkyJq26_97U6S5T2QspoSV5vjf93u8tihcQLCFu4LtdGKIXbrkPXi8BqitU0UfFKWQYLQgC-PHR90lxltIOgBpTQp4WJzjDCDF0Wtysyi4oa6xWpXFDiDpJ7fsyaZ9CLE1eUe9D7K1vy9YNMiRdyuBHKIreBl9aX_ZbXQobf4nb0g2d9s-KJ0a4pJ9P7_Pi5uOHb5efF1fXn9aXq6uFJAz1CwTJRmxoxTCpKaspRAAoBRqqpUIYgAZKA5FEoqkBpUqajcFGKAY3UCqtFT4vXh109y4kPjmSeDaDZc2GgUysD4QKYsf30XYi3vIgLP-7EWLLRW5OOs2JEZhRURuCUVXJmlIJiWCAYNowWeGs9X66bdh0Wh0scDPR-Ym3W96Gn5yiBsCmyQJvJoEYfgw69byz2W7nhNdhGP-7zoNklLCMvv4Hfbi7iWpFbsB6E_K9chTlK5IJmuVQppYPUPlRurN5ltrYvD8reDsryEyvf_etGFLi669f_p-9_j5nL47YrRau36bghjFGaQ5WB1DGkFLU5t5kCPiY_js3-Jh-PqU_l708HtB90V3c8R92Uf9F</recordid><startdate>20210709</startdate><enddate>20210709</enddate><creator>Bearham, Jade</creator><creator>Krutrök, Nina</creator><creator>Lindberg, Botilda</creator><creator>Woodall, Maximillian</creator><creator>Astrand, Annika</creator><creator>Taylor, John D</creator><creator>Biggart, Matthew</creator><creator>Vasiljevs, Stanislavs</creator><creator>Tarran, Robert</creator><creator>Baines, Deborah L</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-7229-0244</orcidid></search><sort><creationdate>20210709</creationdate><title>A modified fluorescent sensor for reporting glucose concentration in the airway lumen</title><author>Bearham, Jade ; Krutrök, Nina ; Lindberg, Botilda ; Woodall, Maximillian ; Astrand, Annika ; Taylor, John D ; Biggart, Matthew ; Vasiljevs, Stanislavs ; Tarran, Robert ; Baines, Deborah L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Analysis</topic><topic>Animals</topic><topic>Biology and Life Sciences</topic><topic>Biosensing Techniques - methods</topic><topic>Bronchus</topic><topic>Calcium-Binding Proteins</topic><topic>Chemical compounds</topic><topic>Dextrose</topic><topic>Diabetes</topic><topic>Diabetes mellitus</topic><topic>E coli</topic><topic>Engineering and Technology</topic><topic>Epithelial cells</topic><topic>Epithelium</topic><topic>Escherichia coli</topic><topic>Escherichia coli - metabolism</topic><topic>Ethylenediamine</topic><topic>Fluorescence</topic><topic>Fluorescent Dyes - chemistry</topic><topic>Fluorophores</topic><topic>Galactose</topic><topic>Genomics</topic><topic>Glucose</topic><topic>Glucose - metabolism</topic><topic>Hemodialysis</topic><topic>Humans</topic><topic>Immunology</topic><topic>Inhalation</topic><topic>Lungs</topic><topic>Measurement</topic><topic>Medicine and Health Sciences</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Molecular structure</topic><topic>Monosaccharide Transport Proteins - metabolism</topic><topic>Mutagenesis</topic><topic>Mutation</topic><topic>Periplasmic Binding Proteins - metabolism</topic><topic>Physical Sciences</topic><topic>Plasmids</topic><topic>Protein expression</topic><topic>Proteins</topic><topic>R&amp;D</topic><topic>Research &amp; development</topic><topic>Research and Analysis Methods</topic><topic>Respiration</topic><topic>Respiratory Mucosa - metabolism</topic><topic>Respiratory tract</topic><topic>Sensors</topic><topic>Staphylococcus infections</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bearham, Jade</creatorcontrib><creatorcontrib>Krutrök, Nina</creatorcontrib><creatorcontrib>Lindberg, Botilda</creatorcontrib><creatorcontrib>Woodall, Maximillian</creatorcontrib><creatorcontrib>Astrand, Annika</creatorcontrib><creatorcontrib>Taylor, John D</creatorcontrib><creatorcontrib>Biggart, Matthew</creatorcontrib><creatorcontrib>Vasiljevs, Stanislavs</creatorcontrib><creatorcontrib>Tarran, Robert</creatorcontrib><creatorcontrib>Baines, Deborah L</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing &amp; Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological &amp; Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>ProQuest Health and Medical</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science &amp; Engineering Collection</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>Advanced Technologies &amp; Aerospace Collection</collection><collection>Agricultural &amp; Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>https://resources.nclive.org/materials</collection><collection>Nursing &amp; Allied Health Database (Alumni Edition)</collection><collection>Meteorological &amp; Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agriculture Science Database</collection><collection>Health &amp; Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>ProQuest Biological Science Journals</collection><collection>Engineering Database</collection><collection>Nursing &amp; Allied Health Premium</collection><collection>ProQuest advanced technologies &amp; aerospace journals</collection><collection>ProQuest Advanced Technologies &amp; Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials science collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bearham, Jade</au><au>Krutrök, Nina</au><au>Lindberg, Botilda</au><au>Woodall, Maximillian</au><au>Astrand, Annika</au><au>Taylor, John D</au><au>Biggart, Matthew</au><au>Vasiljevs, Stanislavs</au><au>Tarran, Robert</au><au>Baines, Deborah L</au><au>Koval, Michael</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A modified fluorescent sensor for reporting glucose concentration in the airway lumen</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2021-07-09</date><risdate>2021</risdate><volume>16</volume><issue>7</issue><spage>e0254248</spage><pages>e0254248-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N'-Dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations &lt;50μM. Using nasal inhalation of the sensor and spectral unmixing to generate images, E149C/A213R GBP-IANBD fluorescence was detected in luminal regions of cryosections of the murine distal lung that was greater in db/db than wildtype mice. In conclusion, this sensor provides a useful tool for further development to measure luminal glucose concentration in models of lung/airway to explore how this may change in disease.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>34242292</pmid><doi>10.1371/journal.pone.0254248</doi><tpages>e0254248</tpages><orcidid>https://orcid.org/0000-0001-7229-0244</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 1932-6203
ispartof PloS one, 2021-07, Vol.16 (7), p.e0254248
issn 1932-6203
1932-6203
language eng
recordid cdi_plos_journals_2549936790
source Publicly Available Content Database; PubMed Central
subjects Analysis
Animals
Biology and Life Sciences
Biosensing Techniques - methods
Bronchus
Calcium-Binding Proteins
Chemical compounds
Dextrose
Diabetes
Diabetes mellitus
E coli
Engineering and Technology
Epithelial cells
Epithelium
Escherichia coli
Escherichia coli - metabolism
Ethylenediamine
Fluorescence
Fluorescent Dyes - chemistry
Fluorophores
Galactose
Genomics
Glucose
Glucose - metabolism
Hemodialysis
Humans
Immunology
Inhalation
Lungs
Measurement
Medicine and Health Sciences
Mice
Mice, Inbred C57BL
Molecular structure
Monosaccharide Transport Proteins - metabolism
Mutagenesis
Mutation
Periplasmic Binding Proteins - metabolism
Physical Sciences
Plasmids
Protein expression
Proteins
R&D
Research & development
Research and Analysis Methods
Respiration
Respiratory Mucosa - metabolism
Respiratory tract
Sensors
Staphylococcus infections
title A modified fluorescent sensor for reporting glucose concentration in the airway lumen
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-01T08%3A35%3A57IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20modified%20fluorescent%20sensor%20for%20reporting%20glucose%20concentration%20in%20the%20airway%20lumen&rft.jtitle=PloS%20one&rft.au=Bearham,%20Jade&rft.date=2021-07-09&rft.volume=16&rft.issue=7&rft.spage=e0254248&rft.pages=e0254248-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0254248&rft_dat=%3Cgale_plos_%3EA667980252%3C/gale_plos_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c692t-216bab84936589581200dd078ecd230071cf12c2a75088dcfbf3fad91b1cdeed3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2549936790&rft_id=info:pmid/34242292&rft_galeid=A667980252&rfr_iscdi=true