Nanoscale binding site localization by molecular distance estimation on native cell surfaces using topological image averaging.

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Tytuł:: Nanoscale binding site localization by molecular distance estimation on native cell surfaces using topological image averaging.
Autorzy:: Kumra Ahnlide V; Division of Infection Medicine, Department of Clinical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
Kumra Ahnlide J; Division of Infection Medicine, Department of Clinical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
Wrighton S; Division of Infection Medicine, Department of Clinical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
Beech JP; Division of Solid State Physics, Department of Physics, Lund University, Lund, Sweden.
Nordenfelt P; Division of Infection Medicine, Department of Clinical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
Źródło:: ELife [Elife] 2022 Feb 24; Vol. 11. Date of Electronic Publication: 2022 Feb 24.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Original Publication: Cambridge, UK : eLife Sciences Publications, Ltd., 2012-
MeSH Terms:: Binding Sites, Antibody*
Antibodies, Monoclonal/*metabolism
Cell Membrane/*metabolism
Epitope Mapping/*methods
Membrane Proteins/*metabolism
Binding Sites ; Cell Membrane/immunology ; Epitopes/metabolism ; Humans ; Ligands ; Membrane Proteins/immunology ; Models, Molecular
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Contributed Indexing:: Keywords: S. pyogenes; antibodies; binding site determination; cell biology; cell surface proteins; epitope; fluorescence microscopy; human; immunology; inflammation; nanometer-scale localization
Local Abstract: [plain-language-summary] Antibodies play a key role in the immune system. These proteins stick to harmful substances, such as bacteria and other disease-causing pathogens, marking them for destruction or blocking their attack. Antibodies are highly selective, and this ability has been used to target particular molecules in research, diagnostics and therapies. Typically, antibodies need to stick to a particular segment, or ‘epitope’, on the surface of a cell in order to trigger an immune response. Knowing where these regions are can help explain how these immune proteins work and aid the development of more effective drugs and diagnostic tools. One way to identify these sites is to measure the nano-distance between antibodies and other features on the cell surface. To do this, researchers take multiple images of the cell the antibody is attached to using light microscopy. Various statistical methods are then applied to create an ‘average image’ that has a higher resolution and can therefore be used to measure the distance between these two points more accurately . While this approach works on fixed shapes, like a perfect circle, it cannot handle human cells and bacteria which are less uniform and have more complex surfaces. Here, Kumra Ahnlide et al. have developed a new method called SiteLoc which can overcome this barrier. The method involves two fluorescent probes: one attached to a specific site on the cell’s surface, and the other to the antibody or another molecule of interest. These two probes emit different colours when imaged with a fluorescent microscope. To cope with objects that have uneven surfaces, such as cells and bacteria, the two signals are transformed to ‘follow’ the same geometrical shape. The relative distance between them is then measured using statistical methods. Using this approach, Kumra Ahnlide et al. were able to identify epitopes on a bacterium, and measure distances on the surface of human red blood cells. The SiteLoc system could make it easier to develop antibody-based treatments and diagnostic tools. Furthermore, it could also be beneficial to the wider research community who could use it to probe other questions that require measuring nanoscale distances.
Molecular Sequence:: Dryad 10.5061/dryad.h18931znn
Substance Nomenclature:: 0 (Antibodies, Monoclonal)
0 (Epitopes)
0 (Ligands)
0 (Membrane Proteins)
Entry Date(s):: Date Created: 20220224 Date Completed: 20220317 Latest Revision: 20220317
Update Code:: 20240104
PubMed Central ID:: PMC8871386
DOI:: 10.7554/eLife.64709
PMID:: 35200140
: Czasopismo naukowe

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Antibody binding to cell surface proteins plays a crucial role in immunity, and the location of an epitope can altogether determine the immunological outcome of a host-target interaction. Techniques available today for epitope identification are costly, time-consuming, and unsuited for high-throughput analysis. Fast and efficient screening of epitope location can be useful for the development of therapeutic monoclonal antibodies and vaccines. Cellular morphology typically varies, and antibodies often bind heterogeneously across a cell surface, making traditional particle-averaging strategies challenging for accurate native antibody localization. In the present work, we have developed a method, SiteLoc, for imaging-based molecular localization on cellular surface proteins. Nanometer-scale resolution is achieved through localization in one dimension, namely, the distance from a bound ligand to a reference surface. This is done by using topological image averaging. Our results show that this method is well suited for antibody binding site measurements on native cell surface morphology and that it can be applied to other molecular distance estimations as well.
Competing Interests: VK, JK, SW, JB, PN No competing interests declared
(© 2022, Kumra Ahnlide et al.)

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