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Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove

Mitogen-activated protein kinases (MAPK) are important regulatory units in cells and they take part in the regulation of many cellular functions such as cell division, differentiation or apoptosis. All MAPKs have a shallow docking groove that interacts with linear binding motifs of their substrate p...

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Autores principales: Alexa, Anita, Ember, Orsolya, Szabó, Ildikó, Mo’ath, Yousef, Póti, Ádám L., Reményi, Attila, Bánóczi, Zoltán
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8281026/
https://www.ncbi.nlm.nih.gov/pubmed/34277705
http://dx.doi.org/10.3389/fmolb.2021.690429
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author Alexa, Anita
Ember, Orsolya
Szabó, Ildikó
Mo’ath, Yousef
Póti, Ádám L.
Reményi, Attila
Bánóczi, Zoltán
author_facet Alexa, Anita
Ember, Orsolya
Szabó, Ildikó
Mo’ath, Yousef
Póti, Ádám L.
Reményi, Attila
Bánóczi, Zoltán
author_sort Alexa, Anita
collection PubMed
description Mitogen-activated protein kinases (MAPK) are important regulatory units in cells and they take part in the regulation of many cellular functions such as cell division, differentiation or apoptosis. All MAPKs have a shallow docking groove that interacts with linear binding motifs of their substrate proteins and their regulatory proteins such as kinases, phosphatases, scaffolds. Inhibition of these protein–protein interactions may reduce or abolish the activity of the targeted kinase. Based on the wide range of their biological activity, this kind of inhibition can be useful in the treatment of many disorders like tumors, inflammation or undesired cell apoptosis. In this study a linear binding motif from the RHDF1 protein—a 15 amino acids long peptide—was selected for optimization to increase its cellular uptake but retaining its low micromolar binding affinity. First, we synthesized an octaarginine conjugate that showed efficient cellular uptake. Next, we set out to reduce the size of this construct. We were able to decrease the length of the original peptide, and to increase its cellular uptake with specific chemical modifications. These new constructs bound better to ERK2 and p38 kinases than the original peptide and they showed markedly increased cellular uptake. The new octaarginine conjugate and one of the minimized bicyclic derivatives could inhibit the phosphorylation of intracellular ERK or p38. However, the modulation of MAPK phosphorylation levels by these cell-penetrating peptides were complex, despite that in biochemical assays they all inhibited MAPK-substrate binding as well as phosphorylation. The optimized peptides depending on the applied concentration caused an expected decrease, but also some unexpected increase in MAPK phosphorylation patterns in the cell. This possibly reflects the complexity of MAPK docking groove mediated protein–protein interactions including bone fide MAPK clients such activator kinases, deactivating phosphatases or regulatory scaffolds. Thus, our findings with optimized cell-penetrating “inhibitory” peptides highlight the opportunities but also the pitfalls of docking peptide based MAPK activity regulation and call for a better quantitative understanding of MAPK mediated protein–protein interactions in cells.
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spelling pubmed-82810262021-07-16 Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove Alexa, Anita Ember, Orsolya Szabó, Ildikó Mo’ath, Yousef Póti, Ádám L. Reményi, Attila Bánóczi, Zoltán Front Mol Biosci Molecular Biosciences Mitogen-activated protein kinases (MAPK) are important regulatory units in cells and they take part in the regulation of many cellular functions such as cell division, differentiation or apoptosis. All MAPKs have a shallow docking groove that interacts with linear binding motifs of their substrate proteins and their regulatory proteins such as kinases, phosphatases, scaffolds. Inhibition of these protein–protein interactions may reduce or abolish the activity of the targeted kinase. Based on the wide range of their biological activity, this kind of inhibition can be useful in the treatment of many disorders like tumors, inflammation or undesired cell apoptosis. In this study a linear binding motif from the RHDF1 protein—a 15 amino acids long peptide—was selected for optimization to increase its cellular uptake but retaining its low micromolar binding affinity. First, we synthesized an octaarginine conjugate that showed efficient cellular uptake. Next, we set out to reduce the size of this construct. We were able to decrease the length of the original peptide, and to increase its cellular uptake with specific chemical modifications. These new constructs bound better to ERK2 and p38 kinases than the original peptide and they showed markedly increased cellular uptake. The new octaarginine conjugate and one of the minimized bicyclic derivatives could inhibit the phosphorylation of intracellular ERK or p38. However, the modulation of MAPK phosphorylation levels by these cell-penetrating peptides were complex, despite that in biochemical assays they all inhibited MAPK-substrate binding as well as phosphorylation. The optimized peptides depending on the applied concentration caused an expected decrease, but also some unexpected increase in MAPK phosphorylation patterns in the cell. This possibly reflects the complexity of MAPK docking groove mediated protein–protein interactions including bone fide MAPK clients such activator kinases, deactivating phosphatases or regulatory scaffolds. Thus, our findings with optimized cell-penetrating “inhibitory” peptides highlight the opportunities but also the pitfalls of docking peptide based MAPK activity regulation and call for a better quantitative understanding of MAPK mediated protein–protein interactions in cells. Frontiers Media S.A. 2021-07-01 /pmc/articles/PMC8281026/ /pubmed/34277705 http://dx.doi.org/10.3389/fmolb.2021.690429 Text en Copyright © 2021 Alexa, Ember, Szabó, Mo’ath, Póti, Reményi and Bánóczi. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Alexa, Anita
Ember, Orsolya
Szabó, Ildikó
Mo’ath, Yousef
Póti, Ádám L.
Reményi, Attila
Bánóczi, Zoltán
Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title_full Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title_fullStr Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title_full_unstemmed Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title_short Peptide Based Inhibitors of Protein Binding to the Mitogen-Activated Protein Kinase Docking Groove
title_sort peptide based inhibitors of protein binding to the mitogen-activated protein kinase docking groove
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8281026/
https://www.ncbi.nlm.nih.gov/pubmed/34277705
http://dx.doi.org/10.3389/fmolb.2021.690429
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