A study of protein–drug interaction based on solvent-induced protein aggregation by fluorescence correlation spectroscopy

Xue, Caining; Yu, Wenxin; Song, Hang; Huang, Xiangyi; Ren, Jicun

doi:10.1039/d2an00031h

Cited by 3 publications

(8 citation statements)

References 42 publications

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“…† One kind of FCS curve (red curve) was similar to the previously reported FCS curves of protein monomers, and we attributed it to the contribution of the EGFP-LC3B monomer only. 30 Another kind of FCS curve (blue curve) shifted significantly to the right compared with the former and exhibited different characteristic from that of the fluorescent monomer and EGFP-LC3B monomer, and we attributed it to the contribution of the EGFP-LC3B punctae and EGFP-LC3B monomer. 30 The absolute values of the fitting residuals were almost less than 0.4 indicating that the two typical kinds of FCS curves can be well fitted.…”

Section: The Monitoring Of Autophagosomes In Living Cellsmentioning

confidence: 73%

“…30 Another kind of FCS curve (blue curve) shifted significantly to the right compared with the former and exhibited different characteristic from that of the fluorescent monomer and EGFP-LC3B monomer, and we attributed it to the contribution of the EGFP-LC3B punctae and EGFP-LC3B monomer. 30 The absolute values of the fitting residuals were almost less than 0.4 indicating that the two typical kinds of FCS curves can be well fitted. As a comparison, we also constructed the HeLa cells, H1299 cells and HEK293T cells stably expressing EGFP-LC3BΔG and EGFP.…”

Section: The Monitoring Of Autophagosomes In Living Cellsmentioning

confidence: 73%

“…Here, a larger τ D value means a larger molecular weight of the diffusing molecules. 30 Brightness per particle (BPP) represents the average brightness of fluorescent molecules within a detection volume, which can be obtained according to eqn (2).…”

Section: Gðτþmentioning

confidence: 99%

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In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

Song

Dong

Ren

2023

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Section: The Monitoring Of Autophagosomes In Living Cellsmentioning

confidence: 73%

Section: The Monitoring Of Autophagosomes In Living Cellsmentioning

confidence: 73%

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In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

Song

Dong

Ren

2023

Analyst

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“…Due to the lack of drugs for diseases caused by protein aggregation related to protein LLPS, identifying biomolecules inhibiting abnormal LLPS and evaluating their inhibitory effects have become important research directions for the development of therapeutic approaches against related protein diseases. Some biomolecules can stabilize protein structures and thus inhibit protein phase separation, such as ATP, which forms ATP clusters on the surface of proteins by binding with highly flexible or hydrophobic regions, thus improving the stability of thermal perturbations. − In our previous studies, we have found FCS to be a reliable research tool for obtaining data on the inhibitory effects of drugs on target proteins. , Therefore, in this section, we tried to evaluate the inhibitory effect of some biomolecules on protein phase separation by FCS.…”

Section: Resultsmentioning

confidence: 99%

“…37−39 In our previous studies, we have found FCS to be a reliable research tool for obtaining data on the inhibitory effects of drugs on target proteins. 40,41 Therefore, in this section, we tried to evaluate the inhibitory effect of some biomolecules on protein phase separation by FCS.…”

Section: Validation Of the Phase Separation Behavior Of Fus-egfp By Fcsmentioning

confidence: 99%

Study on Phase Separation of Fused in Sarcoma by Fluorescence Correlation Spectroscopy

Yu,

Liu,

Huang

et al. 2023

Langmuir

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Liquid−liquid phase separation (LLPS) of fused in sarcoma (FUS) has emerged as a fundamental principle underpinning cellular function and malfunction. However, we know little about the FUS phase transition process from individual molecules to nanoscale condensates, which plays important roles in neurodegenerative diseases. Here, we propose the fluorescence correlation spectroscopy (FCS) method to quantitatively study the phase separation process of FUS protein with the fluorescent tag-enhanced green fluorescent protein (EGFP), from individual molecules to nanoscale condensates. The characteristic diffusion time (τ D ) of the protein condensates can be obtained from the FCS curve, which increases with the growth of the protein hydration radius. The bigger the τ D value of the protein condensates, the larger the condensates formed by the phase separation of FUS. By this method, we discovered that the critical concentration for FUS to phase separation was 20 nM. We then plotted FUS phase diagrams based on τ D under different concentrations of NaCl and found that both low-salt and high-salt concentrations tended to promote FUS-EGFP phase separation. Our results showed that ATP has a good inhibitory effect on FUS phase separation, and its inhibition constant IC 50 was 3.2 mM. Finally, we evaluated the inhibition efficiency of single-stranded DNA sequences (ssDNA) on FUS phase separation and demonstrated that ssDNA containing three copies of TCCCCGT had relatively strong inhibition efficiency. In summary, our work provides detailed insight into the FUS phase transition process from individual molecules to nanoscale condensates at nanomolar concentrations and can be exploited for drug screening of neurodegenerative diseases.

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Co‐Assembled Nanoparticles toward Multi‐Target Combinational Therapy of Alzheimer's Disease by Making Full Use of Molecular Recognition and Self‐Assembly

Li,

Xu,

Wang

et al. 2024

Advanced Materials

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The complex pathologies in Alzheimer's disease (AD) severely limits the effectiveness of single‐target pharmic interventions, thus necessitating multi‐pronged therapeutic strategies. While flexibility is essentially demanded in constructing such multi‐target systems, for achieving optimal synergies and also accommodating the inherent heterogeneity within AD. Utilizing the dynamic reversibility of supramolecular strategy for conferring sufficient tunability in component substitution and proportion adjustment, amphiphilic calixarenes are poised to be a privileged molecular tool for facilely achieving function integration. Herein, taking β‐amyloid (Aβ) fibrillation and oxidative stress as model combination pattern, we proposed a supramolecular multifunctional integration by co‐assembling guanidinium‐modified calixarene with ascorbyl palmitate and loading dipotassium phytate within calixarene cavity. Serial pivotal events can be simultaneously addressed by this versatile system, including (1) inhibition of Aβ production and aggregation, (2) disintegration of Aβ fibrils, (3) acceleration of Aβ metabolic clearance, and (4) regulation of oxidative stress, which is verified to significantly ameliorate the cognitive impairment of 5×FAD mice, with reduced Aβ plaque content, neuroinflammation and neuronal apoptosis. Confronted with the extremely intricate clinical realities of AD, the strategy presented here exhibits ample adaptability for necessary alterations on combinations, thereby may immensely expedite the advancement of AD combinational therapy through providing an exceptionally convenient platform.This article is protected by copyright. All rights reserved

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A study of protein–drug interaction based on solvent-induced protein aggregation by fluorescence correlation spectroscopy

Abstract: Identifying the molecular targets for the beneficial effects of small-molecule drugs is a crucial and currently unsolved challenge which leads to high costs and long development cycles. Therefore, it is...

Cited by 3 publications

References 42 publications

In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

Study on Phase Separation of Fused in Sarcoma by Fluorescence Correlation Spectroscopy

Co‐Assembled Nanoparticles toward Multi‐Target Combinational Therapy of Alzheimer's Disease by Making Full Use of Molecular Recognition and Self‐Assembly

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