Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy

Boassa, Daniela; Lemieux, Sakina P.; Lev‐Ram, Varda; Hu, Junru; Xiong, Qing; Phan, Sébastien; Mackey, Mason; Ramachandra, Ranjan; Peace, Ryan Emily; Adams, Stephen; Ellisman, Mark H.; Ngo, John T.

doi:10.1016/j.chembiol.2019.07.007

Cited by 30 publications

(38 citation statements)

References 30 publications

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“…The target protein must perform several vital functions in the cell, which are disrupted when this protein coaggregates with an effector peptide. To assess possible binding, various approaches can be used both in silico based on interaction patterns [ 213 , 214 ], and in vitro using spectrophotometry, NMR, or electron microscopy [ 215 , 216 , 217 ].…”

Section: Mechanisms Of Antimicrobial Action Of Peptidesmentioning

confidence: 99%

Antimicrobial and Amyloidogenic Activity of Peptides. Can Antimicrobial Peptides Be Used against SARS-CoV-2?

Kurpe

Grishin

Surin

et al. 2020

IJMS

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At present, much attention is paid to the use of antimicrobial peptides (AMPs) of natural and artificial origin to combat pathogens. AMPs have several points that determine their biological activity. We analyzed the structural properties of AMPs, as well as described their mechanism of action and impact on pathogenic bacteria and viruses. Recently published data on the development of new AMP drugs based on a combination of molecular design and genetic engineering approaches are presented. In this article, we have focused on information on the amyloidogenic properties of AMP. This review examines AMP development strategies from the perspective of the current high prevalence of antibiotic-resistant bacteria, and the potential prospects and challenges of using AMPs against infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

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Section: Mechanisms Of Antimicrobial Action Of Peptidesmentioning

confidence: 99%

Antimicrobial and Amyloidogenic Activity of Peptides. Can Antimicrobial Peptides Be Used against SARS-CoV-2?

Kurpe

Grishin

Surin

et al. 2020

IJMS

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“…Singlet oxygen generators (SOG) are known to exert proximity labeling capabilities through lightactivated generation of reactive singlet oxygen species that oxidize biomolecules in nanometer vicinity 28,29 . Small molecule SOG 30 and genetically encodable SOG are extensively used throughout life sciences for photodynamic inactivation of proteins [31][32][33][34] and cells 35,36 , for correlative light-electron microscopy 37 and for the detection of intracellular protein-protein interactions 38,39 in a variety of organisms. The capability of SOG to decode cell surface interactions was recently demonstrated by identifying the binding target of a nerve-highlighting fluorescent peptide in mammalian tissues 40 .…”

Section: Introductionmentioning

confidence: 99%

Light-mediated discovery of surfaceome nanoscale organization and intercellular receptor interaction networks

Müller

Gräbnitz

Barandun

et al. 2020

Preprint

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Delineating the molecular nanoscale organization of the surfaceome is pre-requisite for understanding cellular signaling. Technologies for mapping the spatial relationships of cell surface receptors and their extracellular signaling synapses would open up theranostic opportunities and the possibility to engineer extracellular signaling. Here, we developed an optoproteomic technology termed LUX-MS that exploits singlet oxygen generators (SOG) for the light-triggered identification of acute protein interactions on living cells. Using SOG-coupled antibodies, small molecule-drugs, biologics and intact viral particles, we show that not only ligand-receptor interactions can be decoded across organisms, but also the surfaceome receptor nanoscale organization ligands engage in with direct implications for drug action. Furthermore, investigation of functional immunosynapses revealed that intercellular signaling inbetween APCs and CD8+ T cells can be mapped now providing insights into T cell activation with spatiotemporal resolution. LUX-MS based decoding of surfaceome signaling architectures provides unprecedented molecular insights for the rational development of theranostic strategies.

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“…miniSOG (106 residues) with a noncovalently bound fluorophore flavin mononucleotide (FMN) is a protein photosensitizer with sufficient 1 O 2 quantum yield (≥0.03) [53,54]. Other than generating 1 O 2 , miniSOG, upon excitation with blue light (448 nm), also emits green (500 nm) fluorescence [55,56] and therefore can be used as a fluorescent tag for protein subcellular localization and protein-protein interactions [57][58][59][60][61].…”

Section: Discussionmentioning

confidence: 99%

Permanent Photodynamic Activation of the Cholecystokinin 2 Receptor

Tang

Cui

2020

Biomolecules

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The cholecystokinin 2 receptor (CCK2R) is expressed in the central nervous system and peripheral tissues, playing an important role in higher nervous and gastrointestinal functions, pain sensation, and cancer growth. CCK2R is reversibly activated by cholecystokinin or gastrin, but whether it can be activated permanently is not known. In this work, we found that CCK2R expressed ectopically in CHO-K1 cells was permanently activated in the dark by sulfonated aluminum phthalocyanine (SALPC/AlPcS4, 10–1000 nM), as monitored by Fura-2 fluorescent calcium imaging. Permanent CCK2R activation was also observed with AlPcS2, but not PcS4. CCK2R previously exposed to SALPC (3 and 10 nM) was sensitized by subsequent light irradiation (>580 nm, 31.5 mW·cm−2). After the genetically encoded protein photosensitizer mini singlet oxygen generator (miniSOG) was fused to the N-terminus of CCK2R and expressed in CHO-K1 cells, light irradiation (450 nm, 85 mW·cm−2) activated in-frame CCK2R (miniSOG-CCK2R), permanently triggering persistent calcium oscillations blocked by the CCK2R antagonist YM 022 (30 nM). From these data, it is concluded that SALPC is a long-lasting CCK2R agonist in the dark, and CCK2R is photogenetically activated permanently with miniSOG as photosensitizer. These properties of SALPC and CCK2R could be used to study CCK2R physiology and possibly for pain and cancer therapies.

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Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy

Cited by 30 publications

References 30 publications

Antimicrobial and Amyloidogenic Activity of Peptides. Can Antimicrobial Peptides Be Used against SARS-CoV-2?

Antimicrobial and Amyloidogenic Activity of Peptides. Can Antimicrobial Peptides Be Used against SARS-CoV-2?

Light-mediated discovery of surfaceome nanoscale organization and intercellular receptor interaction networks

Permanent Photodynamic Activation of the Cholecystokinin 2 Receptor

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