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The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)

Most serine proteases are synthesized as inactive zymogens that are activated by cleavage by another protease in a tightly regulated mechanism. The urokinase-type plasminogen activator (uPA) and plasmin cleave and activate each other, constituting a positive feedback loop. How this mutual activation...

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Autores principales: Torres-Paris, Constanza, Chen, Yueyi, Xiao, Lufan, Song, Harriet J., Chen, Pingyu, Komives, Elizabeth A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10520878/
https://www.ncbi.nlm.nih.gov/pubmed/37607618
http://dx.doi.org/10.1016/j.jbc.2023.105179
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author Torres-Paris, Constanza
Chen, Yueyi
Xiao, Lufan
Song, Harriet J.
Chen, Pingyu
Komives, Elizabeth A.
author_facet Torres-Paris, Constanza
Chen, Yueyi
Xiao, Lufan
Song, Harriet J.
Chen, Pingyu
Komives, Elizabeth A.
author_sort Torres-Paris, Constanza
collection PubMed
description Most serine proteases are synthesized as inactive zymogens that are activated by cleavage by another protease in a tightly regulated mechanism. The urokinase-type plasminogen activator (uPA) and plasmin cleave and activate each other, constituting a positive feedback loop. How this mutual activation cycle begins has remained a mystery. We used hydrogen deuterium exchange mass spectrometry to characterize the dynamic differences between the inactive single-chain uPA (scuPA) and its active form two-chain uPA (tcuPA). The results show that the C-terminal β-barrel and the area around the new N terminus have significantly reduced dynamics in tcuPA as compared with scuPA. We also show that the zymogen scuPA is inactive but can, upon storage, become active in the absence of external proteases. In addition to plasmin, the tcuPA can activate scuPA by cleavage at K158, a process called autoactivation. Unexpectedly, tcuPA can cleave at position 158 even when this site is mutated. TcuPA can also cleave scuPA after K135 or K136 in the disordered linker, which generates the soluble protease domain of uPA. Plasmin cleaves scuPA exclusively after K158 and at a faster rate than tcuPA. We propose a mechanism by which the uPA receptor dimerization could promote autoactivation of scuPA on cell surfaces. These results resolve long-standing controversies in the literature surrounding the mechanism of uPA activation.
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spelling pubmed-105208782023-09-27 The autoactivation of human single-chain urokinase-type plasminogen activator (uPA) Torres-Paris, Constanza Chen, Yueyi Xiao, Lufan Song, Harriet J. Chen, Pingyu Komives, Elizabeth A. J Biol Chem Research Article Most serine proteases are synthesized as inactive zymogens that are activated by cleavage by another protease in a tightly regulated mechanism. The urokinase-type plasminogen activator (uPA) and plasmin cleave and activate each other, constituting a positive feedback loop. How this mutual activation cycle begins has remained a mystery. We used hydrogen deuterium exchange mass spectrometry to characterize the dynamic differences between the inactive single-chain uPA (scuPA) and its active form two-chain uPA (tcuPA). The results show that the C-terminal β-barrel and the area around the new N terminus have significantly reduced dynamics in tcuPA as compared with scuPA. We also show that the zymogen scuPA is inactive but can, upon storage, become active in the absence of external proteases. In addition to plasmin, the tcuPA can activate scuPA by cleavage at K158, a process called autoactivation. Unexpectedly, tcuPA can cleave at position 158 even when this site is mutated. TcuPA can also cleave scuPA after K135 or K136 in the disordered linker, which generates the soluble protease domain of uPA. Plasmin cleaves scuPA exclusively after K158 and at a faster rate than tcuPA. We propose a mechanism by which the uPA receptor dimerization could promote autoactivation of scuPA on cell surfaces. These results resolve long-standing controversies in the literature surrounding the mechanism of uPA activation. American Society for Biochemistry and Molecular Biology 2023-08-20 /pmc/articles/PMC10520878/ /pubmed/37607618 http://dx.doi.org/10.1016/j.jbc.2023.105179 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Torres-Paris, Constanza
Chen, Yueyi
Xiao, Lufan
Song, Harriet J.
Chen, Pingyu
Komives, Elizabeth A.
The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title_full The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title_fullStr The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title_full_unstemmed The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title_short The autoactivation of human single-chain urokinase-type plasminogen activator (uPA)
title_sort autoactivation of human single-chain urokinase-type plasminogen activator (upa)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10520878/
https://www.ncbi.nlm.nih.gov/pubmed/37607618
http://dx.doi.org/10.1016/j.jbc.2023.105179
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