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Kathrine Tejlgård Jensen

Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin

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Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin. / Harwood, Seandean Lykke; Nielsen, Nadia Sukusu; Diep, Khang; Jensen, Kathrine Tejlgård; Nielsen, Peter Kresten; Yamamoto, Kazuhiro; Enghild, Jan J.

I: The Journal of Biological Chemistry, Bind 297, Nr. 1, 15.06.2021, s. 100879.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Harwood, Seandean Lykke ; Nielsen, Nadia Sukusu ; Diep, Khang ; Jensen, Kathrine Tejlgård ; Nielsen, Peter Kresten ; Yamamoto, Kazuhiro ; Enghild, Jan J. / Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin. I: The Journal of Biological Chemistry. 2021 ; Bind 297, Nr. 1. s. 100879.

Bibtex

@article{641a705ce15b46479d9a045bea830fd7,
title = "Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin",
abstract = "Human α2-macroglobulin (A2M) is an abundant protease inhibitor in plasma, which regulates many proteolytic processes and is involved in innate immunity. A2M's unique protease-trapping mechanism of inhibition is initiated when a protease cleaves within the exposed and highly susceptible {"}bait region.{"} As the wild-type bait region is permissive to cleavage by most human proteases, A2M is accordingly a broad-spectrum protease inhibitor. In this study, we extensively modified the bait region in order to identify any potential functionally important elements in the bait region sequence and to engineer A2M proteins with restrictive bait regions, which more selectively inhibit a target protease. A2M in which the bait region was entirely replaced by glycine-serine repeats remained fully functional and was not cleaved by any tested protease. Therefore, this bait region was designated as the {"}tabula rasa{"} bait region and used as the starting point for further bait region engineering. Cleavage of the tabula rasa bait region by specific proteases was conveyed by the insertion of appropriate substrate sequences, e.g., basic residues for trypsin. Screening and optimization of tabula rasa bait regions incorporating matrix metalloprotease 2 (MMP2) substrate sequences produced an A2M that was specifically cleaved by MMPs and inhibited MMP2 cleavage activity as efficiently as wild-type A2M. We propose that this approach can be used to develop A2M-based protease inhibitors, which selectively inhibit target proteases, which might be applied toward the clinical inhibition of dysregulated proteolysis as occurs in arthritis and many types of cancer.",
author = "Harwood, {Seandean Lykke} and Nielsen, {Nadia Sukusu} and Khang Diep and Jensen, {Kathrine Tejlg{\aa}rd} and Nielsen, {Peter Kresten} and Kazuhiro Yamamoto and Enghild, {Jan J}",
note = "Copyright {\textcopyright} 2021 The Authors. Published by Elsevier Inc. All rights reserved.",
year = "2021",
month = jun,
day = "15",
doi = "10.1016/j.jbc.2021.100879",
language = "English",
volume = "297",
pages = "100879",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "1",

}

RIS

TY - JOUR

T1 - Development of selective protease inhibitors via engineering of the bait region of human α2-macroglobulin

AU - Harwood, Seandean Lykke

AU - Nielsen, Nadia Sukusu

AU - Diep, Khang

AU - Jensen, Kathrine Tejlgård

AU - Nielsen, Peter Kresten

AU - Yamamoto, Kazuhiro

AU - Enghild, Jan J

N1 - Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.

PY - 2021/6/15

Y1 - 2021/6/15

N2 - Human α2-macroglobulin (A2M) is an abundant protease inhibitor in plasma, which regulates many proteolytic processes and is involved in innate immunity. A2M's unique protease-trapping mechanism of inhibition is initiated when a protease cleaves within the exposed and highly susceptible "bait region." As the wild-type bait region is permissive to cleavage by most human proteases, A2M is accordingly a broad-spectrum protease inhibitor. In this study, we extensively modified the bait region in order to identify any potential functionally important elements in the bait region sequence and to engineer A2M proteins with restrictive bait regions, which more selectively inhibit a target protease. A2M in which the bait region was entirely replaced by glycine-serine repeats remained fully functional and was not cleaved by any tested protease. Therefore, this bait region was designated as the "tabula rasa" bait region and used as the starting point for further bait region engineering. Cleavage of the tabula rasa bait region by specific proteases was conveyed by the insertion of appropriate substrate sequences, e.g., basic residues for trypsin. Screening and optimization of tabula rasa bait regions incorporating matrix metalloprotease 2 (MMP2) substrate sequences produced an A2M that was specifically cleaved by MMPs and inhibited MMP2 cleavage activity as efficiently as wild-type A2M. We propose that this approach can be used to develop A2M-based protease inhibitors, which selectively inhibit target proteases, which might be applied toward the clinical inhibition of dysregulated proteolysis as occurs in arthritis and many types of cancer.

AB - Human α2-macroglobulin (A2M) is an abundant protease inhibitor in plasma, which regulates many proteolytic processes and is involved in innate immunity. A2M's unique protease-trapping mechanism of inhibition is initiated when a protease cleaves within the exposed and highly susceptible "bait region." As the wild-type bait region is permissive to cleavage by most human proteases, A2M is accordingly a broad-spectrum protease inhibitor. In this study, we extensively modified the bait region in order to identify any potential functionally important elements in the bait region sequence and to engineer A2M proteins with restrictive bait regions, which more selectively inhibit a target protease. A2M in which the bait region was entirely replaced by glycine-serine repeats remained fully functional and was not cleaved by any tested protease. Therefore, this bait region was designated as the "tabula rasa" bait region and used as the starting point for further bait region engineering. Cleavage of the tabula rasa bait region by specific proteases was conveyed by the insertion of appropriate substrate sequences, e.g., basic residues for trypsin. Screening and optimization of tabula rasa bait regions incorporating matrix metalloprotease 2 (MMP2) substrate sequences produced an A2M that was specifically cleaved by MMPs and inhibited MMP2 cleavage activity as efficiently as wild-type A2M. We propose that this approach can be used to develop A2M-based protease inhibitors, which selectively inhibit target proteases, which might be applied toward the clinical inhibition of dysregulated proteolysis as occurs in arthritis and many types of cancer.

U2 - 10.1016/j.jbc.2021.100879

DO - 10.1016/j.jbc.2021.100879

M3 - Journal article

C2 - 34139236

VL - 297

SP - 100879

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 1

ER -