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Thromb Haemost 1999; 81(03): 407-414
DOI: 10.1055/s-0037-1614487
Review Article
Schattauer GmbH

Solvent Effects on Activity and Conformation of Plasminogen Activator Inhibitor-1

P. A. Andreasen
1   From the Department of Molecular and Structural Biology, Aarhus University, Denmark
,
R. Egelund
1   From the Department of Molecular and Structural Biology, Aarhus University, Denmark
,
S. Jensen
1   From the Department of Molecular and Structural Biology, Aarhus University, Denmark
,
K. W. Rodenburg
1   From the Department of Molecular and Structural Biology, Aarhus University, Denmark
› Author AffiliationsThis work was supported financially by the Danish Cancer Society, the Danish Heart Foundation, the Danish Medical Research Council, the Danish Biotechnology Programme, Aarhus University Research Foundation, and the NOVO-Nordisk Foundation.
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Publication History

Received22 July 1998

Accepted after resubmission18 November 1998

Publication Date:
09 December 2017 (online)

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Summary

We have studied effects of the solvent composition on the activity and the conformation of human plasminogen activator inhibitor-1 (PAI-1) from HT-1080 fibrosarcoma cells. Non-ionic detergents, including Triton X-100, reduced the inhibitory activity of PAI-1 more than 20-fold at 0° C, but less than 2-fold at 37° C, while glycerol partly prevented the detergent-induced activity-loss at 0° C. The activity-loss was associated with an increase in PAI-1 substrate behaviour. Evaluating the PAI-1 conformation by proteolytic susceptibility of specific peptide bonds, we found that the V8-proteinase susceptibility of the Glu332-Ser333 (P17-P16) bond, part of the hinge between the reactive centre loop (RCL) and β-strand 5A, and the endoproteinase Asp-N susceptibility of several bonds in the β-strand 2A-α-helix E region were increased by detergents at both 0 and 37° C. The susceptibility of the Gln321-Ala322 and the Lys325-Val326 bonds in β-strand 5A to papain and trypsin, respectively, was increased by detergents at 0° C, but not at 37° C, showing a strict correlation between proteinase susceptibility of β-strand 5A and activity-loss at 0° C. Since the β-strand 2A-α-helix E region also showed differential susceptibility to endoproteinase Asp-N in latent, active, and reactive centre-cleaved PAI-1, we propose that a detergent-induced conformational change of the β-strand 2A-α-helix E region influences the movements of β-sheet A, resulting in a cold-induced conformational change of β-strand 5A and thereby an increased substrate behaviour at low temperatures. These results provide new information about the structural basis for serpin substrate behaviour.

 

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