Identification of the cis‑molecular neighbours of the immune checkpoint protein�B7‑H4 in the breast cancer cell‑line SK‑BR‑3 by proteomic proximity labelling

Rees, Johanna S.; Cheung, Lawrence C C; Hamaia, Samir; Davies, G C; Sandercock, Alan M.; Lilley, Kathryn S.; Tigue, Natalie J.; Jackson, Antony P.

doi:10.3892/ijo.2020.5037

Cited by 4 publications

(6 citation statements)

References 34 publications

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“…Peroxidase-based PLPs, such as ascorbate peroxidase (APEX) ( 13 ), Biotinylation by Antibody Recognition (BAR) ( 14 ), Selective Proteomic Proximity Labeling Assay using Tyramide (SPPLAT) ( 15 ), and Enzyme Mediated Activation of Radical Sources (EMARS), have emerged as among the most widely used techniques for interactome mapping and have facilitated significant advances in fundamental understanding across a number of biological fields, including subcellular RNA localization ( 16 ), cell-surface interactomes ( 17 – 19 ), and neurobiology ( 20 ). These platforms rely on horseradish peroxidase (HRP) or APEX enzymes to activate phenol precursors into reactive phenoxy radicals in solution.…”

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confidence: 99%

Radius measurement via super-resolution microscopy enables the development of a variable radii proximity labeling platform

Oakley

Buksh

Fernández

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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The elucidation of protein interaction networks is critical to understanding fundamental biology as well as developing new therapeutics. Proximity labeling platforms (PLPs) are state-of-the-art technologies that enable the discovery and delineation of biomolecular networks through the identification of protein-protein interactions. These platforms work via catalytic generation of reactive probes at a biological region of interest; these probes then diffuse through solution and covalently “tag” proximal biomolecules. The physical distance that the probes diffuse determines the effective labeling radius of the PLP and is a critical parameter that influences the scale and resolution of interactome mapping. As such, by expanding the degrees of labeling resolution offered by PLPs, it is possible to better capture the various size scales of interactomes. At present, however, there is little quantitative understanding of the labeling radii of different PLPs. Here, we report the development of a superresolution microscopy-based assay for the direct quantification of PLP labeling radii. Using this assay, we provide direct extracellular measurements of the labeling radii of state-of-the-art antibody-targeted PLPs, including the peroxidase-based phenoxy radical platform (269 ± 41 nm) and the high-resolution iridium-catalyzed µMap technology (54 ± 12 nm). Last, we apply these insights to the development of a molecular diffusion-based approach to tuning PLP resolution and introduce a new aryl-azide-based µMap platform with an intermediate labeling radius (80 ± 28 nm).

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mentioning

confidence: 99%

Radius measurement via super-resolution microscopy enables the development of a variable radii proximity labeling platform

Oakley

Buksh

Fernández

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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“…Thus, proteins lying within a limited vicinity of the targeted surface protein — here CD3 — are selectively biotinylated. The biotinylated proteins are then enriched through streptavidin affinity pulldown and identified through mass spectrometry [ 18 , 19 , 25 , 26 ]. To our knowledge, this current work is the first use of such a technique in primary cells.…”

Section: Resultsmentioning

confidence: 99%

Proteomic analysis in primary T cells reveals IL-7 alters T cell receptor thresholding via CYTIP/cytohesin/LFA-1 localisation and activation

Queiroz

Piper

Rees

et al. 2022

Biochemical Journal

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The ability of the cellular immune system to discriminate self from foreign antigens depends on the appropriate calibration of the T-cell receptor (TCR) signalling threshold. The lymphocyte homeostatic cytokine interleukin 7 (IL-7) is known to affect TCR thresholding, but the molecular mechanism is not fully elucidated. A better understanding of this process is highly relevant in the context of autoimmune disease therapy and cancer immunotherapy. We sought to characterise the early signalling events attributable to IL-7 priming; in particular, the altered phosphorylation of signal transduction proteins and their molecular localisation to the TCR. By integrating high-resolution proximity- phospho-proteomic and imaging approaches using primary T cells, rather than engineered cell lines or an in vitro expanded T cell population, we uncovered transduction events previously not linked to IL-7. We show that IL-7 leads to dephosphorylation of cytohesin interacting protein (CYTIP) at a hitherto undescribed phosphorylation site (pThr280) and alters the co-localisation of cytohesin 1 with the TCR and LFA-1 integrin. These results show that IL-7, acting via CYTIP and cytohesin-1, may impact TCR activation thresholds by enhancing the co-clustering of TCR and LFA-1 integrin.

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“…Human B7x (hB7x) is on chromosome 1p12/13.1 and inhibits the proliferation of T cells and arrests the cell cycle 82 . In the regulation of immune T cell responses, the immune checkpoint protein B7‐H4 plays a significant part 83 . The binding of T cells from B7‐H4 to CD4 + and CD8 + prevents their activation and proliferation 83 .…”

Section: The Classification Of Immune Checkpointsmentioning

confidence: 99%

“…In the regulation of immune T cell responses, the immune checkpoint protein B7‐H4 plays a significant part 83 . The binding of T cells from B7‐H4 to CD4 + and CD8 + prevents their activation and proliferation 83 . B7‐H4 is predominantly expressed in B cells, macrophages, mature DCs, and T cells 84 .…”

Section: The Classification Of Immune Checkpointsmentioning

confidence: 99%

“…B7‐H4 is predominantly expressed in B cells, macrophages, mature DCs, and T cells 84 . However, the trans‐binding partner in the T cell plasma membrane for B7‐H4 has not been verified yet 83 . With 87% shared amino acids between the human and mice versions, this molecule comprises one IgV and one IgC domain 14 .…”

Section: The Classification Of Immune Checkpointsmentioning

confidence: 99%

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B7 immune checkpoint family members as putative therapeutics in autoimmune disease: An updated overview

et al. 2022

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Autoimmune diseases, especially among young people in the US, are one of the leading causes of morbidity and death. The immune responses are the fundamental pathogenicity of autoimmune disorders. The equilibrium between stimulatory and inhibitory signals is critical for the stimulation, migration, survival, and T cell‐related immune responses. The B7 family can substantially regulate T cell‐mediated immune responses. Nevertheless, recent breakthroughs in immune checkpoint blockade in cancer immunotherapy have facilitated autoimmune diseases, especially among the prone populations. In the current study, we tried to concisely review the role of the B7 family in regulating immune reactions and the influence of immune checkpoint inhibitors on autoimmunity development.

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Identification of the cis‑molecular neighbours of the immune checkpoint protein�B7‑H4 in the breast cancer cell‑line SK‑BR‑3 by proteomic proximity labelling

Cited by 4 publications

References 34 publications

Radius measurement via super-resolution microscopy enables the development of a variable radii proximity labeling platform

Radius measurement via super-resolution microscopy enables the development of a variable radii proximity labeling platform

Proteomic analysis in primary T cells reveals IL-7 alters T cell receptor thresholding via CYTIP/cytohesin/LFA-1 localisation and activation

B7 immune checkpoint family members as putative therapeutics in autoimmune disease: An updated overview

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