Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators
Cell-adhesion molecules (CAMs) are responsible for cell–cell, cell–extracellular matrix, and cell–pathogen interactions. Claudins (CLDNs), occludin (OCLN), and junctional adhesion molecules (JAMs) are CAMs’ components of the tight junction (TJ), the single protein structure tasked with safeguarding...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/12/9784 |
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| author | Jay Rollins Tyler Worthington Allison Dransfield Jordan Whitney Jordan Stanford Emily Hooke Joseph Hobson Jacob Wengler Sandra Hope Dario Mizrachi |
| author_facet | Jay Rollins Tyler Worthington Allison Dransfield Jordan Whitney Jordan Stanford Emily Hooke Joseph Hobson Jacob Wengler Sandra Hope Dario Mizrachi |
| author_sort | Jay Rollins |
| collection | DOAJ |
| description | Cell-adhesion molecules (CAMs) are responsible for cell–cell, cell–extracellular matrix, and cell–pathogen interactions. Claudins (CLDNs), occludin (OCLN), and junctional adhesion molecules (JAMs) are CAMs’ components of the tight junction (TJ), the single protein structure tasked with safeguarding the paracellular space. The TJ is responsible for controlling paracellular permeability according to size and charge. Currently, there are no therapeutic solutions to modulate the TJ. Here, we describe the expression of CLDN proteins in the outer membrane of <i>E. coli</i> and report its consequences. When the expression is induced, the unicellular behavior of <i>E. coli</i> is replaced with multicellular aggregations that can be quantified using Flow Cytometry (FC). Our method, called iCLASP (inspection of cell-adhesion molecules aggregation through FC protocols), allows high-throughput screening (HTS) of small-molecules for interactions with CAMs. Here, we focused on using iCLASP to identify paracellular modulators for CLDN2. Furthermore, we validated those compounds in the mammalian cell line A549 as a proof-of-concept for the iCLASP method. |
| first_indexed | 2024-03-11T02:22:59Z |
| format | Article |
| id | doaj.art-eae0ea2f721443748eee034484fc1702 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T02:22:59Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-eae0ea2f721443748eee034484fc17022023-11-18T10:44:03ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-06-012412978410.3390/ijms24129784Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular ModulatorsJay Rollins0Tyler Worthington1Allison Dransfield2Jordan Whitney3Jordan Stanford4Emily Hooke5Joseph Hobson6Jacob Wengler7Sandra Hope8Dario Mizrachi9Department of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Microbiology and Molecular Biology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USADepartment of Cell Biology and Physiology, College of Life Sciences, Brigham Young University, Provo, UT 84602, USACell-adhesion molecules (CAMs) are responsible for cell–cell, cell–extracellular matrix, and cell–pathogen interactions. Claudins (CLDNs), occludin (OCLN), and junctional adhesion molecules (JAMs) are CAMs’ components of the tight junction (TJ), the single protein structure tasked with safeguarding the paracellular space. The TJ is responsible for controlling paracellular permeability according to size and charge. Currently, there are no therapeutic solutions to modulate the TJ. Here, we describe the expression of CLDN proteins in the outer membrane of <i>E. coli</i> and report its consequences. When the expression is induced, the unicellular behavior of <i>E. coli</i> is replaced with multicellular aggregations that can be quantified using Flow Cytometry (FC). Our method, called iCLASP (inspection of cell-adhesion molecules aggregation through FC protocols), allows high-throughput screening (HTS) of small-molecules for interactions with CAMs. Here, we focused on using iCLASP to identify paracellular modulators for CLDN2. Furthermore, we validated those compounds in the mammalian cell line A549 as a proof-of-concept for the iCLASP method.https://www.mdpi.com/1422-0067/24/12/9784cell-adhesion moleculesparacellular permeabilitytight junctions |
| spellingShingle | Jay Rollins Tyler Worthington Allison Dransfield Jordan Whitney Jordan Stanford Emily Hooke Joseph Hobson Jacob Wengler Sandra Hope Dario Mizrachi Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators International Journal of Molecular Sciences cell-adhesion molecules paracellular permeability tight junctions |
| title | Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators |
| title_full | Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators |
| title_fullStr | Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators |
| title_full_unstemmed | Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators |
| title_short | Expression of Cell-Adhesion Molecules in <i>E. coli</i>: A High Throughput Screening to Identify Paracellular Modulators |
| title_sort | expression of cell adhesion molecules in i e coli i a high throughput screening to identify paracellular modulators |
| topic | cell-adhesion molecules paracellular permeability tight junctions |
| url | https://www.mdpi.com/1422-0067/24/12/9784 |
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