Fragment Screening Yields a Small‐Molecule Stabilizer of 14‐3‐3 Dimers That Modulates Client Protein Interactions

Brink, Hendrik J.; Riemens, Rick; Thee, Stephanie; Bieshuizen, Berend; Pereira, Daniel Da Costa; Esch, Iwan J. P. de; Smit, Martine J.; Boer, Albertus H. de

doi:10.1002/cbic.202200178

Cited by 6 publications

(10 citation statements)

References 38 publications

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“…14-3-3 protein binding can regulate expression of other molecules [ 88 ] and modify interaction networks. [ 8 ] Many 14-3-3 and 14-3-3 PPIs inhibitors [ 89 , 90 , 91 ] and stabilizers [ 92 , 93 , 94 , 95 ] have been regarded as potential therapeutic targets in other systems than heart [ 22 , 23 ]. Small molecules, peptides and natural products that target 14-3-3 or 14-3-3 PPIs have been applied as 14-3-3 and 14-3-3 PPI modulators [ 96 , 97 , 98 ].…”

Section: Discussionmentioning

confidence: 99%

Proteomic Landscape and Deduced Functions of the Cardiac 14-3-3 Protein Interactome

Qu¹,

Tarasov²,

Chakir³

et al. 2022

Cells

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Rationale: The 14-3-3 protein family is known to interact with many proteins in non-cardiac cell types to regulate multiple signaling pathways, particularly those relating to energy and protein homeostasis; and the 14-3-3 network is a therapeutic target of critical metabolic and proteostatic signaling in cancer and neurological diseases. Although the heart is critically sensitive to nutrient and energy alterations, and multiple signaling pathways coordinate to maintain the cardiac cell homeostasis, neither the structure of cardiac 14-3-3 protein interactome, nor potential functional roles of 14-3-3 protein–protein interactions (PPIs) in heart has been explored. Objective: To establish the comprehensive landscape and characterize the functional role of cardiac 14-3-3 PPIs. Methods and Results: We evaluated both RNA expression and protein abundance of 14-3-3 isoforms in mouse heart, followed by co-immunoprecipitation of 14-3-3 proteins and mass spectrometry in left ventricle. We identified 52 proteins comprising the cardiac 14-3-3 interactome. Multiple bioinformatic analyses indicated that more than half of the proteins bound to 14-3-3 are related to mitochondria; and the deduced functions of the mitochondrial 14-3-3 network are to regulate cardiac ATP production via interactions with mitochondrial inner membrane proteins, especially those in mitochondrial complex I. Binding to ribosomal proteins, 14-3-3 proteins likely coordinate protein synthesis and protein quality control. Localizations of 14-3-3 proteins to mitochondria and ribosome were validated via immunofluorescence assays. The deduced function of cardiac 14-3-3 PPIs is to regulate cardiac metabolic homeostasis and proteostasis. Conclusions: Thus, the cardiac 14-3-3 interactome may be a potential therapeutic target in cardiovascular metabolic and proteostatic disease states, as it already is in cancer therapy.

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Section: Discussionmentioning

confidence: 99%

Proteomic Landscape and Deduced Functions of the Cardiac 14-3-3 Protein Interactome

Qu¹,

Tarasov²,

Chakir³

et al. 2022

Cells

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“…Importantly, mutation of these residues has been reported to result in the dissociation of the dimeric form of 14-3-3σ and diminish its function [ 18 , 43 , 44 ]. As the homodimer form is crucial for the full activity of 14-3-3σ, a homodimer stabilizer may be potentially useful in increasing the activity of 14-3-3σ [ 45 , 46 , 47 , 48 ]. A homodimer stabilizer may also be potentially useful to address the downregulation of 14-3-3σ in cancer.…”

Section: Introductionmentioning

confidence: 99%

“…Previously, GCP-based multivalent ligands with a central benzene ring or lysine residues have been reported as a specific stabilizer of 14-3-3ζ/C-Raf and 14-3-3ζ/Tau PPIs [ 54 , 55 ]. Another ligand that was found to be able to stabilize the 14-3-3ζ dimer is fragment 2, but the exact binding site of the ligand on 14-3-3ζ was not resolved experimentally [ 48 ].…”

Section: Introductionmentioning

confidence: 99%

In Silico Studies on GCP-Lys-OMe as a Potential 14-3-3σ Homodimer Stabilizer

Aljabal

Yap²

2022

Pharmaceuticals

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14-3-3 sigma is a vital negative cell cycle regulator. Its expression is consistently downregulated in many types of cancer through gene promoter hypermethylation or proteasomal degradation. 14-3-3 sigma needs to form a homodimer to be functional, while dimers are less prone to degradation than monomers. This suggests that a homodimer stabilizer may increase the tumor suppressive activities of 14-3-3 sigma. However, no known homodimer stabilizer of 14-3-3 sigma has been reported to date. Therefore, this study attempts to test the potential capability of GCP-Lys-OMe (previously reported to bind at the dimer interface of 14-3-3 zeta isoform), to bind and stabilize the 14-3-3 sigma homodimer. In silico docking of GCP-Lys-OMe on 14-3-3 sigma showed more favorable interaction energy (−9.63 kcal/mole) to the dimer interface than 14-3-3 zeta (−7.73 kcal/mole). Subsequent 100 ns molecular dynamics simulation of the GCP-Lys-OMe/14-3-3 sigma complex revealed a highly stable interaction with an average root-mean-square deviation of 0.39 nm (protein backbone) and 0.77 nm (ligand atoms). More contacts between residues at the homodimer interface and a smaller coverage of conformational space of protein atoms were detected for the bound form than for the apo form. These results suggest that GCP-Lys-OMe is a potential homodimer stabilizer of 14-3-3 sigma.

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“…This result suggests that peptide 3 enhanced the binding of the ExoS peptide to the 14-3-3σ protein. This finding is consistent with a previous study on fragment 2, which was also proposed to bind to the dimer interface of the 14-3-3η protein and enhanced the ERα peptide binding to the amphipathic groove of 14-3-3η …”

Section: Resultsmentioning

confidence: 99%

“…This finding is consistent with a previous study on fragment 2, which was also proposed to bind to the dimer interface of the 14-3-3η protein and enhanced the ERα peptide binding to the amphipathic groove of 14-3-3η. 49 Determination of the Enhancement of 14-3-3σ Homodimer Stability by Peptide 3 Using DLS. To further investigate whether peptide 3 is able to stabilize the 14-3-3σ homodimer as predicted in silico, we first tried to understand the intrinsic stability of 14-3-3σ homodimer.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

In Silico Prediction and Biophysical Validation of Novel 14-3-3σ Homodimer Stabilizers

Aljabal,

Teh,

Yap

2023

J. Chem. Inf. Model.

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Fragment Screening Yields a Small‐Molecule Stabilizer of 14‐3‐3 Dimers That Modulates Client Protein Interactions

Cited by 6 publications

References 38 publications

Proteomic Landscape and Deduced Functions of the Cardiac 14-3-3 Protein Interactome

Proteomic Landscape and Deduced Functions of the Cardiac 14-3-3 Protein Interactome

In Silico Studies on GCP-Lys-OMe as a Potential 14-3-3σ Homodimer Stabilizer

In Silico Prediction and Biophysical Validation of Novel 14-3-3σ Homodimer Stabilizers

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