Molecular evolution of plasminogen activator inhibitor-1 functional stability.


Abstract

Plasminogen activator inhibitor-1 (PAI-1) is a member of the serine protease inhibitor (serpin) supergene family and a central regulatory protein in the blood coagulation system. PAI-1 is unique among serpins in exhibiting distinct active and inactive (latent) conformations in vivo. Though the structure of latent PAI-1 was recently solved, the structure of the short-lived, active form of PAI-1 is not known. In order to probe the structural basis for this unique conformational change, a randomly mutated recombinant PAI-1 expression library was constructed in bacteriophage and screened for increased functional stability. Fourteen unique clones were selected, and shown to exhibit functional half-lives (T1/2S) exceeding that of wild-type PAI-1 by up to 72-fold. The most stable variant (T1/2 = 145 h) contained four mutations. Detailed analysis of these four mutations, individually and in combination, demonstrated that the markedly enhanced functional stability of the parent compound mutant required contributions from all four substitutions, with no individual T1/2 exceeding 6.6 h. The functional stability of at least eight of the remaining 13 compound mutants also required interactions between two or more amino acid substitutions, with no single variant increasing the T1/2 by > 10-fold. The nature of the identified mutations implies that the unique instability of the PAI-1 active conformation evolved through global changes in protein packing and suggest a selective advantage for transient inhibitor function.

Submission Details

ID: jwTQqwyw

Submitter: Shu-Ching Ou

Submission Date: Nov. 21, 2018, 11:55 a.m.

Version: 1

Publication Details
Berkenpas MB;Lawrence DA;Ginsburg D,EMBO J (1995) Molecular evolution of plasminogen activator inhibitor-1 functional stability. PMID:7621813
Additional Information

Study Summary

Number of data points 62
Proteins Plasminogen activator inhibitor 1
Unique complexes 31
Assays/Quantities/Protocols Experimental Assay: Half-Life (37°C)
Libraries Mutants of PAI-1

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1A7C 1998-03-12T00:00:00+0000 1.95 HUMAN PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 IN COMPLEX WITH A PENTAPEPTIDE
1B3K 1998-12-11T00:00:00+0000 2.99 Plasminogen activator inhibitor-1
1C5G 1999-12-07T00:00:00+0000 2.6 PLASMINOGEN ACTIVATOR INHIBITOR-1
1DB2 1999-11-02T00:00:00+0000 2.7 CRYSTAL STRUCTURE OF NATIVE PLASMINOGEN ACTIVATOR INHIBITOR-1
1DVM 2000-01-21T00:00:00+0000 2.4 ACTIVE FORM OF HUMAN PAI-1
1DVN 2000-01-21T00:00:00+0000 2.1 LATENT FORM OF PLASMINOGEN ACTIVATOR INHIBITOR-1 (PAI-1)
1LJ5 2002-04-19T00:00:00+0000 1.8 1.8A Resolution Structure of Latent Plasminogen Activator Inhibitor-1(PAI-1)
1OC0 2003-02-03T00:00:00+0000 2.28 plasminogen activator inhibitor-1 complex with somatomedin B domain of vitronectin
3CVM 2008-04-18T00:00:00+0000 2.02 High resolution structure of a stable Plasminogen activator inhibitor type-1 in its protease cleaved form
3EOX 2008-09-29T00:00:00+0000 2.61 High quality structure of cleaved PAI-1-stab

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Plasminogen activator inhibitor 1 P05121 PAI1_HUMAN