Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator

Abstract Although trypsin-like serine proteases have flexible surface-exposed loops and are known to adopt higher and lower activity conformations, structural determinants for the different conformations have remained largely obscure. The trypsin-like serine protease, urokinase-type plasminogen acti...

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Autores principales:	Tobias Kromann-Hansen, Eva Louise Lange, Hans Peter Sørensen, Gholamreza Hassanzadeh-Ghassabeh, Mingdong Huang, Jan K. Jensen, Serge Muyldermans, Paul J. Declerck, Elizabeth A. Komives, Peter A. Andreasen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/409897e4334a4c2c81d516f9a73f9d64
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spelling	oai:doaj.org-article:409897e4334a4c2c81d516f9a73f9d642021-12-02T11:40:33ZDiscovery of a novel conformational equilibrium in urokinase-type plasminogen activator10.1038/s41598-017-03457-72045-2322https://doaj.org/article/409897e4334a4c2c81d516f9a73f9d642017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03457-7https://doaj.org/toc/2045-2322Abstract Although trypsin-like serine proteases have flexible surface-exposed loops and are known to adopt higher and lower activity conformations, structural determinants for the different conformations have remained largely obscure. The trypsin-like serine protease, urokinase-type plasminogen activator (uPA), is central in tissue remodeling processes and also strongly implicated in tumor metastasis. We solved five X-ray crystal structures of murine uPA (muPA) in the absence and presence of allosteric molecules and/or substrate-like molecules. The structure of unbound muPA revealed an unsuspected non-chymotrypsin-like protease conformation in which two β-strands in the core of the protease domain undergoes a major antiparallel-to-parallel conformational transition. We next isolated two anti-muPA nanobodies; an active-site binding nanobody and an allosteric nanobody. Crystal structures of the muPA:nanobody complexes and hydrogen-deuterium exchange mass spectrometry revealed molecular insights about molecular factors controlling the antiparallel-to-parallel equilibrium in muPA. Together with muPA activity assays, the data provide valuable insights into regulatory mechanisms and conformational flexibility of uPA and trypsin-like serine proteases in general.Tobias Kromann-HansenEva Louise LangeHans Peter SørensenGholamreza Hassanzadeh-GhassabehMingdong HuangJan K. JensenSerge MuyldermansPaul J. DeclerckElizabeth A. KomivesPeter A. AndreasenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tobias Kromann-Hansen Eva Louise Lange Hans Peter Sørensen Gholamreza Hassanzadeh-Ghassabeh Mingdong Huang Jan K. Jensen Serge Muyldermans Paul J. Declerck Elizabeth A. Komives Peter A. Andreasen Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
description	Abstract Although trypsin-like serine proteases have flexible surface-exposed loops and are known to adopt higher and lower activity conformations, structural determinants for the different conformations have remained largely obscure. The trypsin-like serine protease, urokinase-type plasminogen activator (uPA), is central in tissue remodeling processes and also strongly implicated in tumor metastasis. We solved five X-ray crystal structures of murine uPA (muPA) in the absence and presence of allosteric molecules and/or substrate-like molecules. The structure of unbound muPA revealed an unsuspected non-chymotrypsin-like protease conformation in which two β-strands in the core of the protease domain undergoes a major antiparallel-to-parallel conformational transition. We next isolated two anti-muPA nanobodies; an active-site binding nanobody and an allosteric nanobody. Crystal structures of the muPA:nanobody complexes and hydrogen-deuterium exchange mass spectrometry revealed molecular insights about molecular factors controlling the antiparallel-to-parallel equilibrium in muPA. Together with muPA activity assays, the data provide valuable insights into regulatory mechanisms and conformational flexibility of uPA and trypsin-like serine proteases in general.
format	article
author	Tobias Kromann-Hansen Eva Louise Lange Hans Peter Sørensen Gholamreza Hassanzadeh-Ghassabeh Mingdong Huang Jan K. Jensen Serge Muyldermans Paul J. Declerck Elizabeth A. Komives Peter A. Andreasen
author_facet	Tobias Kromann-Hansen Eva Louise Lange Hans Peter Sørensen Gholamreza Hassanzadeh-Ghassabeh Mingdong Huang Jan K. Jensen Serge Muyldermans Paul J. Declerck Elizabeth A. Komives Peter A. Andreasen
author_sort	Tobias Kromann-Hansen
title	Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
title_short	Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
title_full	Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
title_fullStr	Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
title_full_unstemmed	Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
title_sort	discovery of a novel conformational equilibrium in urokinase-type plasminogen activator
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/409897e4334a4c2c81d516f9a73f9d64
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_version_	1718395576876269568

Discovery of a novel conformational equilibrium in urokinase-type plasminogen activator

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