Use of Native-PAGE for the Identification of Epichaperomes in Cell Lines

Roychowdhury, Tanaya; Santhaseela, Anand R.; Sharma, Shivani; Panchal, Palak; Rodina, Anna; Chiosis, Gabriela

doi:10.1007/978-1-0716-3342-7_14

Cited by 2 publications

(1 citation statement)

References 20 publications

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“…In our pursuit of understanding epichaperome biology, our team has developed a range of chemical tools and methodologies. These include a radiolabeled [ 124 I]-PU-AD probe for epichaperome detection by positron emission tomography (PET) in mice and humans [ 20 , 23 ], a size-based native Western blot method for epichaperome detection in cell or tissue homogenates [ 8 , 24 ], and a chemoproteomics method termed epichaperomics or dysfunctional Protein–Protein Interactome (dfPPI), which identifies proteins and functions impacted by epichaperomes, allowing for the delineation of processes affected by epichaperome formation in disease [ 8 , 11 ]. However, while these tools have significantly contributed to our understanding of epichaperomes in disease biology, they often lack the requisite sensitivity, cellular resolution, and versatility necessary for addressing fundamental questions pertaining to epichaperome dynamics and composition.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and Characterization of Click Chemical Probes for Single-Cell Resolution Detection of Epichaperomes in Neurodegenerative Disorders

Bay,

Digwal,

Rodilla Martín

et al. 2024

Biomedicines

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Neurodegenerative disorders, including Alzheimer’s disease (AD) and Parkinson’s disease (PD), represent debilitating conditions with complex, poorly understood pathologies. Epichaperomes, pathologic protein assemblies nucleated on key chaperones, have emerged as critical players in the molecular dysfunction underlying these disorders. In this study, we introduce the synthesis and characterization of clickable epichaperome probes, PU-TCO, positive control, and PU-NTCO, negative control. Through comprehensive in vitro assays and cell-based investigations, we establish the specificity of the PU-TCO probe for epichaperomes. Furthermore, we demonstrate the efficacy of PU-TCO in detecting epichaperomes in brain tissue with a cellular resolution, underscoring its potential as a valuable tool for dissecting single-cell responses in neurodegenerative diseases. This clickable probe is therefore poised to address a critical need in the field, offering unprecedented precision and versatility in studying epichaperomes and opening avenues for novel insights into their role in disease pathology.

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Section: Introductionmentioning

confidence: 99%

Synthesis and Characterization of Click Chemical Probes for Single-Cell Resolution Detection of Epichaperomes in Neurodegenerative Disorders

Bay,

Digwal,

Rodilla Martín

et al. 2024

Biomedicines

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Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation

Roychowdhury,

McNutt,

Pasala

et al. 2024

Nat Commun

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The intricate network of protein-chaperone interactions is crucial for maintaining cellular function. Recent discoveries have unveiled the existence of specialized chaperone assemblies, known as epichaperomes, which serve as scaffolding platforms that orchestrate the reconfiguration of protein-protein interaction networks, thereby enhancing cellular adaptability and proliferation. This study explores the structural and regulatory aspects of epichaperomes, with a particular focus on the role of post-translational modifications (PTMs) in their formation and function. A key finding is the identification of specific PTMs on HSP90, particularly at residues Ser226 and Ser255 within an intrinsically disordered region, as critical determinants of epichaperome assembly. Our data demonstrate that phosphorylation of these serine residues enhances HSP90’s interactions with other chaperones and co-chaperones, creating a microenvironment conducive to epichaperome formation. Moreover, we establish a direct link between epichaperome function and cellular physiology, particularly in contexts where robust proliferation and adaptive behavior are essential, such as in cancer and pluripotent stem cell maintenance. These findings not only provide mechanistic insights but also hold promise for the development of novel therapeutic strategies targeting chaperone assemblies in diseases characterized by epichaperome dysregulation, thereby bridging the gap between fundamental research and precision medicine.

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Use of Native-PAGE for the Identification of Epichaperomes in Cell Lines

Cited by 2 publications

References 20 publications

Synthesis and Characterization of Click Chemical Probes for Single-Cell Resolution Detection of Epichaperomes in Neurodegenerative Disorders

Synthesis and Characterization of Click Chemical Probes for Single-Cell Resolution Detection of Epichaperomes in Neurodegenerative Disorders

Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation

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