A KLK4 proteinase substrate capture approach to antagonize PAR1

Abstract Proteinase-activated receptor-1 (PAR1), triggered by thrombin and other serine proteinases such as tissue kallikrein-4 (KLK4), is a key driver of inflammation, tumor invasiveness and tumor metastasis. The PAR1 transmembrane G-protein-coupled receptor therefore represents an attractive targe...

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Autores principales:	Eitan Rabinovitch, Koishiro Mihara, Amiram Sananes, Marianna Zaretsky, Michael Heyne, Julia Shifman, Amir Aharoni, Morley D. Hollenberg, Niv Papo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1a0212dd91364948b979eba1d92f904e
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spelling	oai:doaj.org-article:1a0212dd91364948b979eba1d92f904e2021-12-02T18:50:57ZA KLK4 proteinase substrate capture approach to antagonize PAR110.1038/s41598-021-95666-42045-2322https://doaj.org/article/1a0212dd91364948b979eba1d92f904e2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95666-4https://doaj.org/toc/2045-2322Abstract Proteinase-activated receptor-1 (PAR1), triggered by thrombin and other serine proteinases such as tissue kallikrein-4 (KLK4), is a key driver of inflammation, tumor invasiveness and tumor metastasis. The PAR1 transmembrane G-protein-coupled receptor therefore represents an attractive target for therapeutic inhibitors. We thus used a computational design to develop a new PAR1 antagonist, namely, a catalytically inactive human KLK4 that acts as a proteinase substrate-capture reagent, preventing receptor cleavage (and hence activation) by binding to and occluding the extracellular R41-S42 canonical PAR1 proteolytic activation site. On the basis of in silico site-saturation mutagenesis, we then generated KLK4S207A,L185D, a first-of-a-kind ‘decoy’ PAR1 inhibitor, by mutating the S207A and L185D residues in wild-type KLK4, which strongly binds to PAR1. KLK4S207A,L185D markedly inhibited PAR1 cleavage, and PAR1-mediated MAPK/ERK activation as well as the migration and invasiveness of melanoma cells. This ‘substrate-capturing’ KLK4 variant, engineered to bind to PAR1, illustrates proof of principle for the utility of a KLK4 ‘proteinase substrate capture’ approach to regulate proteinase-mediated PAR1 signaling.Eitan RabinovitchKoishiro MiharaAmiram SananesMarianna ZaretskyMichael HeyneJulia ShifmanAmir AharoniMorley D. HollenbergNiv PapoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eitan Rabinovitch Koishiro Mihara Amiram Sananes Marianna Zaretsky Michael Heyne Julia Shifman Amir Aharoni Morley D. Hollenberg Niv Papo A KLK4 proteinase substrate capture approach to antagonize PAR1
description	Abstract Proteinase-activated receptor-1 (PAR1), triggered by thrombin and other serine proteinases such as tissue kallikrein-4 (KLK4), is a key driver of inflammation, tumor invasiveness and tumor metastasis. The PAR1 transmembrane G-protein-coupled receptor therefore represents an attractive target for therapeutic inhibitors. We thus used a computational design to develop a new PAR1 antagonist, namely, a catalytically inactive human KLK4 that acts as a proteinase substrate-capture reagent, preventing receptor cleavage (and hence activation) by binding to and occluding the extracellular R41-S42 canonical PAR1 proteolytic activation site. On the basis of in silico site-saturation mutagenesis, we then generated KLK4S207A,L185D, a first-of-a-kind ‘decoy’ PAR1 inhibitor, by mutating the S207A and L185D residues in wild-type KLK4, which strongly binds to PAR1. KLK4S207A,L185D markedly inhibited PAR1 cleavage, and PAR1-mediated MAPK/ERK activation as well as the migration and invasiveness of melanoma cells. This ‘substrate-capturing’ KLK4 variant, engineered to bind to PAR1, illustrates proof of principle for the utility of a KLK4 ‘proteinase substrate capture’ approach to regulate proteinase-mediated PAR1 signaling.
format	article
author	Eitan Rabinovitch Koishiro Mihara Amiram Sananes Marianna Zaretsky Michael Heyne Julia Shifman Amir Aharoni Morley D. Hollenberg Niv Papo
author_facet	Eitan Rabinovitch Koishiro Mihara Amiram Sananes Marianna Zaretsky Michael Heyne Julia Shifman Amir Aharoni Morley D. Hollenberg Niv Papo
author_sort	Eitan Rabinovitch
title	A KLK4 proteinase substrate capture approach to antagonize PAR1
title_short	A KLK4 proteinase substrate capture approach to antagonize PAR1
title_full	A KLK4 proteinase substrate capture approach to antagonize PAR1
title_fullStr	A KLK4 proteinase substrate capture approach to antagonize PAR1
title_full_unstemmed	A KLK4 proteinase substrate capture approach to antagonize PAR1
title_sort	klk4 proteinase substrate capture approach to antagonize par1
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1a0212dd91364948b979eba1d92f904e
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A KLK4 proteinase substrate capture approach to antagonize PAR1

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