α1-Antitrypsin Portland, a bioengineered serpin highly selective for furin: Application as an antipathogenic agent

François Jean, Kori Stella, Laurel Thomas, Gseping Liu, Yang Kevin Xiang, Andrew J. Reason, Gary Thomas

Research output: Contribution to journalArticle

221 Citations (Scopus)

Abstract

The important role of furin in the proteolytic activation of many pathogenic molecules has made this endoprotease a target for the development of potent and selective antiproteolytic agents. Here, we demonstrate the utility of the protein-based inhibitor α1-antitrypsin Portland (α1-PDX) as an antipathogenic agent that can be used prophylactically to block furin- dependent cell killing by Pseudomonas exotoxin A. Biochemical analysis of the specificity of a bacterially expressed Hisand FLAG-tagged α1-PDX (α1- PDX/hf) revealed the selectivity of the α1-PDX/hf reactive site loop for furin (K(i), 600 pM) but not for other proprotein convertase family members or other unrelated endoproteases. Kinetic studies show that α1-PDX/hf inhibits furin by a slow tight-binding mechanism characteristic of serpin molecules and functions as a suicide substrate inhibitor. Once bound to furin's active site, α1-PDX/hf partitions with equal probability to undergo proteolysis by furin at the C-terminal side of the reactive center - Arg355-Ile-Pro-Arg358-↓ or to form a kinetically trapped SDS-stable complex with the enzyme. This partitioning between the complex-forming and proteolytic pathways contributes to the ability of α1-PDX/hf to differentially inhibit members of the proprotein convertase family. Finally, we propose a structural model of the α1-PDX-reactive site loop that explains the high degree of enzyme selectivity of this serpin and which can be used to generate small molecule furin inhibitors.

Original languageEnglish (US)
Pages (from-to)7293-7298
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number13
DOIs
StatePublished - Jun 23 1998
Externally publishedYes

Fingerprint

Furin
Serpins
Proprotein Convertases
Catalytic Domain
Aptitude
Structural Models
Enzymes
Suicide
Proteolysis

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

α1-Antitrypsin Portland, a bioengineered serpin highly selective for furin : Application as an antipathogenic agent. / Jean, François; Stella, Kori; Thomas, Laurel; Liu, Gseping; Xiang, Yang Kevin; Reason, Andrew J.; Thomas, Gary.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, No. 13, 23.06.1998, p. 7293-7298.

Research output: Contribution to journalArticle

Jean, François ; Stella, Kori ; Thomas, Laurel ; Liu, Gseping ; Xiang, Yang Kevin ; Reason, Andrew J. ; Thomas, Gary. / α1-Antitrypsin Portland, a bioengineered serpin highly selective for furin : Application as an antipathogenic agent. In: Proceedings of the National Academy of Sciences of the United States of America. 1998 ; Vol. 95, No. 13. pp. 7293-7298.
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