Strategies for Conditional Regulation of Proteins

Nadendla, Karthik; Simpson, Grant G.; Becher, Julie; Journeaux, Toby; Cabeza-Cabrerizo, Mar; Bernardes, Gonçalo J. L.

doi:10.1021/jacsau.2c00654

Cited by 2 publications

(2 citation statements)

References 202 publications

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“…Light-controlled optogenetic tools are intriguing for dynamic regulation , due to their precise control over protein–protein interactions in a spatiotemporal and reversible manner. As optogenetic molecules are relatively large in molecular weights and prone to aggregation upon overexpression in prokaryotic cells, fusion expression of optogenetic molecules with scaffolding and client proteins often leads to the formation of insoluble and inactive aggregates, ,,, which adversely affects the functioning of the resulting condensates and the realization of reversible and repeatable regulation. To address this challenge, here, we propose a modular framework for the organization of synthetic condensates by decoupling condensate formation and client recruitment/release (Scheme ).…”

Section: Introductionmentioning

confidence: 99%

Spatiotemporal Organization of Functional Cargoes by Light-Switchable Condensation in Escherichia coli Cells

Pan,

Zu,

Zhu

et al. 2024

JACS Au

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Biomolecular condensates are dynamic subcellular compartments that lack surrounding membranes and can spatiotemporally organize the cellular biochemistry of eukaryotic cells. However, such dynamic organization has not been realized in prokaryotes that naturally lack organelles, and strategies are urgently needed for dynamic biomolecular compartmentalization. Here we develop a light-switchable condensate system for on-demand dynamic organization of functional cargoes in the model prokaryotic Escherichia coli cells. The condensate system consists of two modularly designed and genetically encoded fusions that contain a condensation-enabling scaffold and a functional cargo fused to the blue light-responsive heterodimerization pair, iLID and SspB, respectively. By appropriately controlling the biogenesis of the protein fusions, the condensate system allows rapid recruitment and release of cargo proteins within seconds in response to light, and this process is also reversible and repeatable. Finally, the system is demonstrated to dynamically control the subcellular localization of a cell division inhibitor, SulA, which enables the reversible regulation of cell morphologies. Therefore, this study provides a new strategy to dynamically control cellular processes by harnessing light-controlled condensates in prokaryotic cells.

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Section: Introductionmentioning

confidence: 99%

Spatiotemporal Organization of Functional Cargoes by Light-Switchable Condensation in Escherichia coli Cells

Pan,

Zu,

Zhu

et al. 2024

JACS Au

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show abstract

“…7 Activatable or conditional therapeutics are a promising therapeutic modality to enhance the efficacy or the safety profile of small-molecule drugs and biologics. [8][9][10][11] Specific triggers (e.g. pH, hypoxia, redox, reactive oxygen species, and enzymes) at the diseased sites are exploited to release or convert pro-therapeutics into their active forms.…”

Section: Introductionmentioning

confidence: 99%

Nanobody-targeted conditional antimicrobial therapeutics

Ngambenjawong,

Ko,

Pishesha

et al. 2024

Preprint

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Conditional therapeutics that rely on disease microenvironment-specific triggers for activation are a promising strategy to improve therapeutic cargos. Among the investigated triggers, protease activity is used most often, because of its dysregulation in several diseases. How to optimally fine-tune protease activation for different therapeutic cargos remains a challenge. Here, we designed nanobody-targeted conditional antimicrobial therapeutics to deliver a model therapeutic peptide and protein to the site of bacterial infection. We explored several parameters that influence proteolytic activation. We report the use of targeting nanobodies to enhance the activation of therapeutics that are otherwise activated inefficiently, despite extensive optimization of the cleavable linker. Specifically, pairing of Ly6G/C or ADAM10-targeting nanobodies with ADAM10-cleavable linkers improved activation via proximity-enabled reactivity. More broadly, this optimization framework provides a guideline for the development of conditional therapeutics to treat various diseases where protease activity is dysregulated.

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Strategies for Conditional Regulation of Proteins

Cited by 2 publications

References 202 publications

Spatiotemporal Organization of Functional Cargoes by Light-Switchable Condensation in Escherichia coli Cells

Spatiotemporal Organization of Functional Cargoes by Light-Switchable Condensation in Escherichia coli Cells

Nanobody-targeted conditional antimicrobial therapeutics

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