Electrochemical biosensors based on peptide-kinase interactions at the kinase docking site

Joshi, Pralhad Namdev; Mervinetsky, Evgeniy; Solomon, Ohad; Chen, Yu‐Ju; Yitzchaik, Shlomo; Friedler, Assaf

doi:10.1016/j.bios.2022.114177

Cited by 13 publications

(14 citation statements)

References 54 publications

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“…[59] To conclude, we showed that inhibition of ERK2 can be done by using peptides that target the PPI of its DRS docking site. [29] This approach is highly versatile and can easily be utilized for different kinases and other types of enzymes, whose activities are crucial in disease.…”

Section: Discussionmentioning

confidence: 99%

“…Using the previously developed electrochemical assay, we evaluated the ability of the peptides to inhibit the interactions of ERK2 with the surface-bound ELK1 D domain peptide, which are mediated by the DRS docking site. [29] The most active peptides penetrated HT29 colon cancer cells and decreased their viability.…”

Section: Figure 2 (A)mentioning

confidence: 98%

“…[27,28] We have recently used the interaction of ERK2 at its DRS docking site for sensing the kinase. [29] The ELK1 D domain peptide, ELK1 312-321, was adsorbed onto gold electrodes to form a peptide layer. Exposure to ERK2 resulted in increased resistance to charge transfer, and surface chemistry analysis showed that ERK2 bound the ELK1 peptide layer, which resulted in the electrochemical sensing.…”

Section: Figure 2 (A)mentioning

confidence: 99%

“…To determine whether the D domain peptides that bound ERK2 can also inhibit its interactions mediated via its DRS, we used an electrochemical impedance spectroscopy (EIS)based ERK2 detection assay developed in our labs. [29] The assay was comprised of the ELK1 D domain peptide, ELK1 312-321, adsorbed onto gold electrodes. To increase the sensitivity of this assay, the ELK1 peptide was co-adsorbed with 1-hexanethiol.…”

Section: The D Domain Peptides Inhibit Erk2 Binding To the Surface-bo...mentioning

confidence: 99%

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Peptide‐Based Inhibitors that Target the Docking Site of ERK2

Solomon

Shpilt

Sapir

et al. 2022

Israel Journal of Chemistry

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Abnormal kinase activity is highly associated with disease, especially cancer. Thus, kinases are important targets for developing anti‐cancer drugs. The common approach for kinase inhibition is using small molecules that compete with ATP for binding the ATP‐binding site of the kinase. However, since the ATP‐binding site is in many cases common to numerous kinases, it is difficult to achieve selectivity when targeting this site. Here we present an alternative approach of targeting the protein‐protein interactions of kinases as means for achieving selectivity in their inhibition. We demonstrate this approach by using peptides for inhibiting the docking D‐recruitment site (DRS) of the kinase ERK2. We designed a library of peptides, derived from DRS binding sequences of ERK2‐binding proteins. We synthesized the peptides and quantified their interactions with ERK2. Three peptides, derived from the proteins ELK1, SAP1 and SAP2, bound ERK2 in the low micromolar range. These peptides also inhibited the interaction of ERK2 with a surface‐bound ELK1 derived peptide. The peptides penetrated HT29 colon cancer cells and induced a moderate decrease in cell viability. Our approach can be further utilized for developing selective peptide‐based kinase inhibitors.

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Section: Discussionmentioning

confidence: 99%

Section: Figure 2 (A)mentioning

confidence: 98%

Section: Figure 2 (A)mentioning

confidence: 99%

Section: The D Domain Peptides Inhibit Erk2 Binding To the Surface-bo...mentioning

confidence: 99%

See 2 more Smart Citations

Peptide‐Based Inhibitors that Target the Docking Site of ERK2

Solomon

Shpilt

Sapir

et al. 2022

Israel Journal of Chemistry

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“…Changes in the arrangement of the peptide layers upon exposure to the desired kinases resulted in their electrochemical detection. Some of these methods are based on the kinase-mediated phosphorylation for sensing, , while the others are based on the protein–protein interactions of the catalytic and docking sites of kinases with their substrates. Several platforms for detecting multiple disease biomarkers were developed .…”

mentioning

confidence: 99%

Using a Single Peptide to Electrochemically Sense Multiple Kinases

et al. 2022

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Kinases are responsible for regulating cellular and physiological processes, and abnormal kinase activity is associated with various diseases. Therefore, kinases are being used as biomarkers for disease and developing methods for their sensing is highly important. Usually more than one kinase is involved in phosphorylating a target protein. However, kinase detection methods usually detect the activity of only one specific kinase. Here we describe an electrochemical kinase sensing tool for the selective detection of two kinases using the same target peptide. We demonstrate the sensing of kinases ERK2 and PKCδ. This is based on a single sensing element, a peptide that contains two distinct phosphorylation sites of these two kinases. Reversibility experiments with alkaline phosphatase and reaction with the electrochemically active ferrocene-labeled ATP showed that the mechanism of sensing is by detecting the enzymatic phosphorylation. Our approach can be further utilized to develop devices for the detection of multiple kinases and can be expanded to other types of enzymes involved in disease.

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Breaking barriers in electrochemical biosensing using bioinspired peptide and phage probes

Campuzano,

Pedrero,

Barderas

et al. 2024

Anal Bioanal Chem

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Electrochemical biosensing continues to advance tirelessly, overcoming barriers that have kept it from leaving research laboratories for many years. Among them, its compromised performance in complex biological matrices due to fouling or receptor stability issues, the limitations in determining toxic and small analytes, and its use, conditioned to the commercial availability of commercial receptors and the exploration of natural molecular interactions, deserved to be highlighted. To address these challenges, in addition to the intrinsic properties of electrochemical biosensing, its coupling with biomimetic materials has played a fundamental role, among which bioinspired phage and peptide probes stand out. The versatility in design and employment of these probes has opened an unimaginable plethora of possibilities for electrochemical biosensing, improving their performance far beyond the development of highly sensitive and selective devices. The state of the art offers robust electroanalytical biotools, capable of operating in complex samples and with exciting opportunities to discover and determine targets regardless of their toxicity and size, the commercial availability of bioreceptors, and prior knowledge of molecular interactions. With all this in mind, this review offers a panoramic, novel, and updated vision of both the tremendous advances and opportunities offered by the combination of electrochemical biosensors with bioinspired phage and peptide probes and the challenges and research efforts that are envisioned in the immediate future. Graphical Abstract

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Electrochemical biosensors based on peptide-kinase interactions at the kinase docking site

Cited by 13 publications

References 54 publications

Peptide‐Based Inhibitors that Target the Docking Site of ERK2

Peptide‐Based Inhibitors that Target the Docking Site of ERK2

Using a Single Peptide to Electrochemically Sense Multiple Kinases

Breaking barriers in electrochemical biosensing using bioinspired peptide and phage probes

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