Umeå University's logo

umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Conformational studies of plasminogen activator inhibitor type 1 by fluorescence spectroscopy: Analysis of the reactive centre of inhibitory and substrate forms, and of their respective reactive-centre cleaved forms
Umeå University, Faculty of Medicine, Medical Biochemistry and Biophsyics.
Faculty of Science and Technology, Chemistry.
Faculty of Science and Technology, Chemistry.
Umeå University, Faculty of Medicine, Medical Biochemistry and Biophsyics.
2000 (English)In: European Journal of Biochemistry, Vol. 267, no 12, p. 3729-34Article in journal (Refereed) Published
Abstract [en]

The inhibitors that belong to the serpin family are suicide inhibitors that control the major proteolytic cascades in eucaryotes. Recent data suggest that serpin inhibition involves reactive centre cleavage followed by loop insertion, whereby the covalently linked protease is translocated away from the initial docking site. However under certain circumstances, serpins can also be cleaved like a substrate by target proteases. In this report we have studied the conformation of the reactive centre of plasminogen activator inhibitor type 1 (PAI-1) mutants with inhibitory and substrate properties. The polarized steady-state and time-resolved fluorescence anisotropies were determined for BODIPY® probes attached to the P1' and P3 positions of the substrate and active forms of PAI-1. The fluorescence data suggest an extended orientational freedom of the probe in the reactive centre of the substrate form as compared to the active form, revealing that the conformation of the reactive centres differ. The intramolecular distance between the P1' and P3 residues in reactive centre cleaved inhibitory and substrate mutants of PAI-1, were determined by using the donor-donor energy migration (DDEM) method. The distances found were 57 ± 4 Å and 63 ± 3 Å, respectively, which is comparable to the distance obtained between the same residues when PAI-1 is in complex with urokinase-type plasminogen activator (uPA). Following reactive centre cleavage, our data suggest that the core of the inhibitory and substrate forms possesses an inherited ability of fully inserting the reactive centre loop into β-sheet A. In the inhibitory forms of PAI-1 forming serpin-protease complexes, this ability leads to a translocation of the cognate protease from one pole of the inhibitor to the opposite one.

Place, publisher, year, edition, pages
2000. Vol. 267, no 12, p. 3729-34
Identifiers
URN: urn:nbn:se:umu:diva-8616DOI: doi:10.1046/j.1432-1327.2000.01406.xOAI: oai:DiVA.org:umu-8616DiVA, id: diva2:148287
Available from: 2008-01-31 Created: 2008-01-31 Last updated: 2018-06-09Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full text

Authority records

Johansson, Lennart B-ÅNy, Tor

Search in DiVA

By author/editor
Johansson, Lennart B-ÅNy, Tor
By organisation
Medical Biochemistry and BiophsyicsChemistry

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 256 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf