Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors

Trine Tamberg, Zebin Hong, Daphné De Schepper, Signe Skovbjerg, Daniel M Dupont, Lars Vitved, Christine R Schar, Karsten Skjoedt, Lotte K Vogel, Jan K Jensen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

10 Downloads (Pure)

Resumé

Matriptase is a member of the type-II transmembrane serine protease (TTSP) family and plays a crucial role in the development and maintenance of epithelial tissues. As all chymotrypsin-like serine proteases, matriptase is synthesized as a zymogen (proform), requiring a cleavage event for full activity. Recent studies suggest that the zymogen of matriptase possesses enough catalytic activity to not only facilitate autoactivation, but also carry out its in vivo functions, which include activating several proteolytic and signaling cascades. Inhibition of zymogen matriptase may therefore be a highly effective approach for limiting matriptase activity. To this end, here we sought to characterize the catalytic activity of human zymogen matriptase and to develop mAb inhibitors against this enzyme form. Using a mutated variant of matriptase in which the serine protease domain is locked in the zymogen conformation, we confirmed that the zymogen form of human matriptase has catalytic activity. Moreover, the crystal structure of the catalytic domain of zymogen matriptase was solved to 2.5 Å resolution to characterize specific antibody-based matriptase inhibitors and to further structure-based studies. Finally, we describe the first antibody-based competitive inhibitors that target both the zymogen and activated forms of matriptase. We propose that these antibodies provide a more efficient way to regulate matriptase activity by targeting the protease both before and after its activation and may be of value for both research and preclinical applications.

OriginalsprogEngelsk
TidsskriftJournal of Biological Chemistry
Vol/bind294
Udgave nummer1
Sider (fra-til)314-326
ISSN0021-9258
DOI
StatusUdgivet - 4. jan. 2019

Fingeraftryk

Enzyme Precursors
Antibodies
Serine Proteases
Catalyst activity
matriptase
Chymotrypsin
Enzyme Inhibitors
Human Activities
Conformations
Catalytic Domain
Peptide Hydrolases
Epithelium
Crystal structure
Chemical activation
Maintenance

Bibliografisk note

© 2019 Tamberg et al.

Citer dette

Tamberg, T., Hong, Z., De Schepper, D., Skovbjerg, S., Dupont, D. M., Vitved, L., ... Jensen, J. K. (2019). Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors. Journal of Biological Chemistry, 294(1), 314-326. https://doi.org/10.1074/jbc.RA118.004126
Tamberg, Trine ; Hong, Zebin ; De Schepper, Daphné ; Skovbjerg, Signe ; Dupont, Daniel M ; Vitved, Lars ; Schar, Christine R ; Skjoedt, Karsten ; Vogel, Lotte K ; Jensen, Jan K. / Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors. I: Journal of Biological Chemistry. 2019 ; Bind 294, Nr. 1. s. 314-326.
@article{c62973cc182249b9b4ab5a7c791c6058,
title = "Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors",
abstract = "Matriptase is a member of the type-II transmembrane serine protease (TTSP) family and plays a crucial role in the development and maintenance of epithelial tissues. As all chymotrypsin-like serine proteases, matriptase is synthesized as a zymogen (proform), requiring a cleavage event for full activity. Recent studies suggest that the zymogen of matriptase possesses enough catalytic activity to not only facilitate autoactivation, but also carry out its in vivo functions, which include activating several proteolytic and signaling cascades. Inhibition of zymogen matriptase may therefore be a highly effective approach for limiting matriptase activity. To this end, here we sought to characterize the catalytic activity of human zymogen matriptase and to develop mAb inhibitors against this enzyme form. Using a mutated variant of matriptase in which the serine protease domain is locked in the zymogen conformation, we confirmed that the zymogen form of human matriptase has catalytic activity. Moreover, the crystal structure of the catalytic domain of zymogen matriptase was solved to 2.5 {\AA} resolution to characterize specific antibody-based matriptase inhibitors and to further structure-based studies. Finally, we describe the first antibody-based competitive inhibitors that target both the zymogen and activated forms of matriptase. We propose that these antibodies provide a more efficient way to regulate matriptase activity by targeting the protease both before and after its activation and may be of value for both research and preclinical applications.",
author = "Trine Tamberg and Zebin Hong and {De Schepper}, Daphn{\'e} and Signe Skovbjerg and Dupont, {Daniel M} and Lars Vitved and Schar, {Christine R} and Karsten Skjoedt and Vogel, {Lotte K} and Jensen, {Jan K}",
note = "{\circledC} 2019 Tamberg et al.",
year = "2019",
month = "1",
day = "4",
doi = "10.1074/jbc.RA118.004126",
language = "English",
volume = "294",
pages = "314--326",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "1",

}

Tamberg, T, Hong, Z, De Schepper, D, Skovbjerg, S, Dupont, DM, Vitved, L, Schar, CR, Skjoedt, K, Vogel, LK & Jensen, JK 2019, 'Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors', Journal of Biological Chemistry, bind 294, nr. 1, s. 314-326. https://doi.org/10.1074/jbc.RA118.004126

Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors. / Tamberg, Trine; Hong, Zebin; De Schepper, Daphné; Skovbjerg, Signe; Dupont, Daniel M; Vitved, Lars; Schar, Christine R; Skjoedt, Karsten; Vogel, Lotte K; Jensen, Jan K.

I: Journal of Biological Chemistry, Bind 294, Nr. 1, 04.01.2019, s. 314-326.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Blocking the proteolytic activity of zymogen matriptase with antibody-based inhibitors

AU - Tamberg, Trine

AU - Hong, Zebin

AU - De Schepper, Daphné

AU - Skovbjerg, Signe

AU - Dupont, Daniel M

AU - Vitved, Lars

AU - Schar, Christine R

AU - Skjoedt, Karsten

AU - Vogel, Lotte K

AU - Jensen, Jan K

N1 - © 2019 Tamberg et al.

PY - 2019/1/4

Y1 - 2019/1/4

N2 - Matriptase is a member of the type-II transmembrane serine protease (TTSP) family and plays a crucial role in the development and maintenance of epithelial tissues. As all chymotrypsin-like serine proteases, matriptase is synthesized as a zymogen (proform), requiring a cleavage event for full activity. Recent studies suggest that the zymogen of matriptase possesses enough catalytic activity to not only facilitate autoactivation, but also carry out its in vivo functions, which include activating several proteolytic and signaling cascades. Inhibition of zymogen matriptase may therefore be a highly effective approach for limiting matriptase activity. To this end, here we sought to characterize the catalytic activity of human zymogen matriptase and to develop mAb inhibitors against this enzyme form. Using a mutated variant of matriptase in which the serine protease domain is locked in the zymogen conformation, we confirmed that the zymogen form of human matriptase has catalytic activity. Moreover, the crystal structure of the catalytic domain of zymogen matriptase was solved to 2.5 Å resolution to characterize specific antibody-based matriptase inhibitors and to further structure-based studies. Finally, we describe the first antibody-based competitive inhibitors that target both the zymogen and activated forms of matriptase. We propose that these antibodies provide a more efficient way to regulate matriptase activity by targeting the protease both before and after its activation and may be of value for both research and preclinical applications.

AB - Matriptase is a member of the type-II transmembrane serine protease (TTSP) family and plays a crucial role in the development and maintenance of epithelial tissues. As all chymotrypsin-like serine proteases, matriptase is synthesized as a zymogen (proform), requiring a cleavage event for full activity. Recent studies suggest that the zymogen of matriptase possesses enough catalytic activity to not only facilitate autoactivation, but also carry out its in vivo functions, which include activating several proteolytic and signaling cascades. Inhibition of zymogen matriptase may therefore be a highly effective approach for limiting matriptase activity. To this end, here we sought to characterize the catalytic activity of human zymogen matriptase and to develop mAb inhibitors against this enzyme form. Using a mutated variant of matriptase in which the serine protease domain is locked in the zymogen conformation, we confirmed that the zymogen form of human matriptase has catalytic activity. Moreover, the crystal structure of the catalytic domain of zymogen matriptase was solved to 2.5 Å resolution to characterize specific antibody-based matriptase inhibitors and to further structure-based studies. Finally, we describe the first antibody-based competitive inhibitors that target both the zymogen and activated forms of matriptase. We propose that these antibodies provide a more efficient way to regulate matriptase activity by targeting the protease both before and after its activation and may be of value for both research and preclinical applications.

U2 - 10.1074/jbc.RA118.004126

DO - 10.1074/jbc.RA118.004126

M3 - Journal article

C2 - 30409910

VL - 294

SP - 314

EP - 326

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 1

ER -