scholarly journals Single fluorescence probes along the reactive center loop reveal site-specific changes during the latency transition of PAI-1

2015 ◽  
Vol 25 (2) ◽  
pp. 487-498
Author(s):  
Tihami Qureshi ◽  
Cynthia B. Peterson
Keyword(s):  
Fluorescence Probes ◽  
Site Specific ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Pai 1

The Reactive-center Loop of Active PAI-1 is Folded Close to the Protein Core and can be Partially Inserted

2004 ◽  
Vol 335 (3) ◽  
pp. 823-832 ◽  
Author(s):  
Peter Hägglöf ◽  
Fredrik Bergström ◽  
Malgorzata Wilczynska ◽  
Lennart B.-Å Johansson ◽  
Tor Ny
Keyword(s):  
Protein Core ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Pai 1

scholarly journals Distinct encounter complexes of PAI-1 with plasminogen activators and vitronectin revealed by changes in the conformation and dynamics of the reactive center loop

2015 ◽  
Vol 25 (2) ◽  
pp. 499-510 ◽  
Author(s):  
Tihami Qureshi ◽  
Sumit Goswami ◽  
Carlee S. McClintock ◽  
Matthew T. Ramsey ◽  
Cynthia B. Peterson
Keyword(s):  
Plasminogen Activators ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Pai 1

scholarly journals The Omptins of Yersinia pestis and Salmonella enterica Cleave the Reactive Center Loop of Plasminogen Activator Inhibitor 1

2010 ◽  
Vol 192 (18) ◽  
pp. 4553-4561 ◽  
Author(s):  
Johanna Haiko ◽  
Liisa Laakkonen ◽  
Katri Juuti ◽  
Nisse Kalkkinen ◽  
Timo K. Korhonen
Keyword(s):  
Yersinia Pestis ◽  
Plasminogen Activator ◽  
Salmonella Enterica ◽  
Yersinia Pseudotuberculosis ◽  
Plasminogen Activator Inhibitor ◽  
Plasminogen Activator Inhibitor 1 ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Activator Inhibitor ◽  
Pai 1

ABSTRACT Plasminogen activator inhibitor 1 (PAI-1) is a serine protease inhibitor (serpin) and a key molecule that regulates fibrinolysis by inactivating human plasminogen activators. Here we show that two important human pathogens, the plague bacterium Yersinia pestis and the enteropathogen Salmonella enterica serovar Typhimurium, inactivate PAI-1 by cleaving the R346-M347 bait peptide bond in the reactive center loop. No cleavage of PAI-1 was detected with Yersinia pseudotuberculosis, an oral/fecal pathogen from which Y. pestis has evolved, or with Escherichia coli. The cleavage and inactivation of PAI-1 were mediated by the outer membrane proteases plasminogen activator Pla of Y. pestis and PgtE protease of S. enterica, which belong to the omptin family of transmembrane endopeptidases identified in Gram-negative bacteria. Cleavage of PAI-1 was also detected with the omptins Epo of Erwinia pyrifoliae and Kop of Klebsiella pneumoniae, which both belong to the same omptin subfamily as Pla and PgtE, whereas no cleavage of PAI-1 was detected with omptins of Shigella flexneri or E. coli or the Yersinia chromosomal omptins, which belong to other omptin subfamilies. The results reveal a novel serpinolytic mechanism by which enterobacterial species expressing omptins of the Pla subfamily bypass normal control of host proteolysis.

scholarly journals Serpin reactive center loop mobility is required for inhibitor function but not for enzyme recognition.

1994 ◽  
Vol 269 (44) ◽  
pp. 27657-27662 ◽  
Author(s):  
D A Lawrence ◽  
S T Olson ◽  
S Palaniappan ◽  
D Ginsburg
Keyword(s):  
Reactive Center Loop ◽  
Reactive Center ◽  
Enzyme Recognition

A reactive center loop–based prediction platform to enhance the design of therapeutic SERPINs

2021 ◽  
Vol 118 (45) ◽  
pp. e2108458118
Author(s):  
Wariya Sanrattana ◽  
Thibaud Sefiane ◽  
Simone Smits ◽  
Nadine D. van Kleef ◽  
Marcel H. Fens ◽  
...  
Keyword(s):  
Cleavage Site ◽  
Serine Proteases ◽  
Protein C ◽  
Activated Protein C ◽  
Reactive Center Loop ◽  
Physiological Processes ◽  
Naturally Occurring ◽  
Reactive Center ◽  
Insight Into

Serine proteases are essential for many physiological processes and require tight regulation by serine protease inhibitors (SERPINs). A disturbed SERPIN–protease balance may result in disease. The reactive center loop (RCL) contains an enzymatic cleavage site between the P1 through P1’ residues that controls SERPIN specificity. This RCL can be modified to improve SERPIN function; however, a lack of insight into sequence–function relationships limits SERPIN development. This is complicated by more than 25 billion mutants needed to screen the entire P4 to P4’ region. Here, we developed a platform to predict the effects of RCL mutagenesis by using α1-antitrypsin as a model SERPIN. We generated variants for each of the residues in P4 to P4’ region, mutating them into each of the 20 naturally occurring amino acids. Subsequently, we profiled the reactivity of the resulting 160 variants against seven proteases involved in coagulation. These profiles formed the basis of an in silico prediction platform for SERPIN inhibitory behavior with combined P4 to P4’ RCL mutations, which were validated experimentally. This prediction platform accurately predicted SERPIN behavior against five out of the seven screened proteases, one of which was activated protein C (APC). Using these findings, a next-generation APC-inhibiting α1-antitrypsin variant was designed (KMPR/RIRA; / indicates the cleavage site). This variant attenuates blood loss in an in vivo hemophilia A model at a lower dosage than the previously developed variant AIKR/KIPP because of improved potency and specificity. We propose that this SERPIN-based RCL mutagenesis approach improves our understanding of SERPIN behavior and will facilitate the design of therapeutic SERPINs.

scholarly journals Crystal Structure of Monomeric Native Antithrombin Reveals a Novel Reactive Center Loop Conformation

2006 ◽  
Vol 281 (46) ◽  
pp. 35478-35486 ◽  
Author(s):  
Daniel J. D. Johnson ◽  
Jonathan Langdown ◽  
Wei Li ◽  
Stephan A. Luis ◽  
Trevor P. Baglin ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Loop Conformation

The Bβ-sheet in the PAI-1 Molecule Plays an Important Role for Its Stability

2000 ◽  
Vol 83 (06) ◽  
pp. 896-901 ◽  
Author(s):  
Guang-Chao Sui ◽  
Björn Wiman
Keyword(s):  
Amino Acids ◽  
Half Life ◽  
Ph Dependence ◽  
The Other ◽  
Wild Type ◽  
E Coli ◽  
Reactive Center ◽  
The Stability ◽  
Β Sheet ◽  
Pai 1

SummaryWe have investigated the B β-sheet in PAI-1 regarding its role for the stability of the molecule. The residues from His219 to Tyr241 (except for Gly230 and Pro240), covering the s2B and s3B strands, and in addition His185 and His190 were substituted by amino acids with opposite properties. The 23 generated single-site changed mutants and also wild type PAI-1 (wtPAI-1) were expressed in E. coli. Subsequently they were purified by heparin-Sepharose and anhydrotrypsin agarose affinity chromatographies. The stability of the purified PAI-1 variants was analyzed at 37° C and at different pHs (5.5, 6.5 or 7.5). At pH 7.5 and 37° C, single substitutions of the residues in the central portions of both strands 2 and 3 in the B β-sheet (Ile223 to Leu226 on s2B and Met235 to Ile237 on s3B), caused a significant decrease in stability, yielding half-lives of about 10–25% as compared to wtPAI-1. On the other hand, mutations at both sides of the central portion of the B β-sheet (Tyr221, Asp222, Tyr228 and Thr232) frequently resulted in an increased PAI-1 stability (up to 7-fold). While wtPAI-1 exhibited prolonged half-lives at pH 6.5 and 5.5, the PAI-1 variant Y228S was more stable at neutral pH (half-life of 9.6 h at pH 7.5) as compared to its half-life at pH 5.5 (1.1 h). One of the 4 modified histidine residues (His229) resulted in a variant with a clearly affected stability as a function of pH, suggesting that it may, at least in part, be of importance for the pH dependence of the PAI-1 stability. Thus, our data demonstrate that the B β-sheet is of great importance for the stability of the molecule. Modifications in this part causes decreased or increased stability in a certain pattern, suggesting effects on the insertion rate of the reactive center loop into the A β-sheet of the molecule.

scholarly journals Pyrexia and acidosis act independently of neutrophil elastase reactive center loop cleavage to effect cortisol release from corticosteroid‐binding globulin

2020 ◽  
Vol 29 (12) ◽  
pp. 2495-2509
Author(s):  
Emily J. Meyer ◽  
David J. Torpy ◽  
Anastasia Chernykh ◽  
Morten Thaysen‐Andersen ◽  
Marni A. Nenke ◽  
...  
Keyword(s):  
Neutrophil Elastase ◽  
Reactive Center Loop ◽  
Reactive Center ◽  
Corticosteroid Binding Globulin ◽  
Cortisol Release
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