scholarly journals Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

2020 ◽  
Author(s):  
Yankel Chekli ◽  
Caroline Peron-Cane ◽  
Dario Dell’Arciprete ◽  
Jean-François Allemand ◽  
Chenge Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Surface Proteins ◽  
Cellular Functions ◽  
Reporter Protein ◽  
Cell Surface Proteins ◽  
Fluorescent Reporter ◽  
Bacterial Proteins

AbstractBacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localization of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and specifically tagged on the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study of the organization and dynamics of the bacterial cell surface proteins.

scholarly journals Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yankel Chekli ◽  
Caroline Peron-Cane ◽  
Dario Dell’Arciprete ◽  
Jean-François Allemand ◽  
Chenge Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Cell Surface ◽  
Bacterial Cell ◽  
Surface Proteins ◽  
Cellular Functions ◽  
Reporter Protein ◽  
Cell Surface Proteins ◽  
Fluorescent Reporter ◽  
Bacterial Proteins

Abstract Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localisation of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and used to specifically tag the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study the organization and dynamics of the bacterial cell surface proteins.

Assessing Changes in the Expression Levels of Cell Surface Proteins with a Turn-on Fluorescent Molecular Probe

2021 ◽  
Author(s):  
Joydev Hatai ◽  
Pragati Kishore Prasad ◽  
Naama Mankovski ◽  
Noa Oppenheimer ◽  
Tamar Unger ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Cell Surface ◽  
Outer Membrane Protein ◽  
Protein C ◽  
Nitrilotriacetic Acid ◽  
Surface Proteins ◽  
Molecular Probe ◽  
Cell Surface Proteins ◽  
Turn On

Tri-nitrilotriacetic acid (NTA)-based fluorescent probes were developed and used to image His-tagged-labelled outer membrane protein C (His-OmpC) in live Escherichia coli. One of these probes was designed to light up...

Nucleic acid-based molecular computation heads towards cellular applications

2021 ◽  
Author(s):  
Lanlan Chen ◽  
Wanzhen Chen ◽  
Guo Liu ◽  
Jingying Li ◽  
Chunhua Lu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Cell Surface ◽  
Surface Proteins ◽  
Specific Interactions ◽  
Cellular Functions ◽  
Cell Surface Proteins ◽  
Molecular Computation ◽  
Cellular Behaviors

Nucleic acid-based molecular computation for cellular applications, including specific interactions with cell surface proteins, biosensing, mimicking cellular behaviors, and engineering cellular functions.

scholarly journals Restricted Mobility of Cell Surface Proteins in the Polar Regions of Escherichia coli

2004 ◽  
Vol 186 (9) ◽  
pp. 2594-2602 ◽  
Author(s):  
Miguel A. de Pedro ◽  
Christoph G. Grünfelder ◽  
Heinz Schwarz
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Cell Envelope ◽  
Envelope Proteins ◽  
Surface Proteins ◽  
Polar Regions ◽  
Free Medium ◽  
Cell Surface Proteins ◽  
Restricted Mobility ◽  
Murein Sacculus

ABSTRACT The polar regions of the Escherichia coli murein sacculus are metabolically inert and stable in time. Because the sacculus and the outer membrane are tightly associated, we investigated whether polar inert murein could restrict the mobility of other cell envelope elements. Cells were covalently labeled with a fluorescent reagent, chased in dye-free medium, and observed by microscopy. Fluorescent material was more efficiently retained at the cell poles than at any other location. The boundary between high and low fluorescence intensity areas was rather sharp. Labeled material consisted mostly of cell envelope proteins, among them the free and murein-bound forms of Braun's lipoprotein. Our results indicate that the mobility of at least some cell envelope proteins is restrained at regions in correspondence with underlying areas of inert murein.

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