Nature 355, 270 - 273 (16 January 1992); doi:10.1038/355270a0

Structural basis of latency in plasminogen activator inhibitor-1

James Mottonen^*, Arne Strand^*, Jindrich Symersky^*, Robert M. Sweet^†, Dennis E. Danley^‡, Kieran F. Geoghegan^‡, Robert D. Gerard^*§ & Elizabeth J. Goldsmith^*

Departments of ^* Biochemistry and ^§ Internal Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75235-9050, USA
^† Department of Biology, Brookhaven National Laboratory, Upton, New York 11973, USA
^‡ Department of Molecular Genetics and Protein Chemistry, Central Research Division, Pfizer Inc., Eastern Point Road, Groton, Connecticut, 06340, USA
To whom correspondence should be addressed.

HUMAN plasminogen activator inhibitor-1 (PAI-1)^1,2 is the fast-acting inhibitor of tissue plasminogen activator and urokinase³ and is a member of the serpin family of protease inhibitors⁴. Serpins normally form complexes with their target proteases that dissociate very slowly as cleaved species and then fold into a highly stable inactive state⁵ in which the residues that flank the scissile bond (P1 and P1'; ref. 6) are separated by about 70 Å (refs 7–9). PAI-1 also spontaneously folds into a stable¹⁰ inactive state without cleavage; this state is termed 'latent' because inhibitory activity can be restored through denaturation and renaturation^2,10. Here we report the structure of intact latent PAI-1 determined by single-crystal X-ray diffraction to 2.6 Å resolution. The three-dimensional structure reveals that residues on the N-terminal side of the primary recognition site are inserted as a central strand of the largest sheet, in positions similar to the corresponding residues in the cleaved form of the serpin ₁proteinase inhibitor (₁-PI)⁷. Residues C-terminal to the recognition site occupy positions on the surface of the molecule distinct from those of the corresponding residues in cleaved serpins^7–9 or in the intact inactive serpin homologue, ovalbumin¹¹, and its cleavage product, plakal-bumin¹². The structure of latent PAI-1 is similar to one formed after cleavage in other serpins, and the stability of both latent PAI-1 and cleaved serpins may be derived from the same structural features^13,14.

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