Antiprotease targeting: altered specificity of α1-antitrypsin by amino acid replacement at the reactive centre

S. Jallat, L. H. Tessier, A. Benavente, Ronald Crystal, M. Courtney

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Alpha-1 antitrypsin (α1AT) is an efficient inhibitor of the human neutrophil proteases, elastase and cathepsin G. The reactive centre P1 residue (Met358) of α1AT is important in defining the specificity of inhibition; furthermore, oxidation of this residue results in a loss of inhibitor activity. There is evidence that oxidative inactivation of α1AT may be involved in the pathogenesis of pulmonary emphysema associated with cigarette smoking. We have studied the effect of a series of amino acid replacements at the active centre on the inhibition properties of α1AT. The mutant proteins were produced in E. coli following in vitro mutagenesis of the α1AT cDNA. Alpha-1-AT (Ile358), (Ala358) and (Val558) were efficient inhibitors of both neutrophil and pancreatic elastase, but not cathepsin G. Alpha-1-AT (Ala356, Val358) and α1AT (Phe358) were specific for pancreatic elastase and cathepsin G respectively. Alpha-1-AT (Leu358) inhibited both neutrophil elastase and cathepsin G. These data show that, for effective inhibition, a potential cleavage site for the protease must be displayed at the α1AT active centre. In each case, replacement of Met358 led to resistance to oxidative inactivation. Since α1AT (Leu358) inhibits both neutrophil proteases and is resistant to oxidation, this variant may be of increased potential for the therapy of destructive lung disorders.

Original languageEnglish
Pages (from-to)287-298
Number of pages12
JournalRevue Francaise de Transfusion et Immuno-Hematologie
Volume29
Issue number4
DOIs
Publication statusPublished - 1 Jan 1986
Externally publishedYes

Fingerprint

Cathepsin G
Protease Inhibitors
Leukocyte Elastase
Pancreatic Elastase
Amino Acids
Peptide Hydrolases
Oxidation
Pulmonary Emphysema
alpha 1-Antitrypsin
Mutagenesis
Mutant Proteins
Tobacco Products
Escherichia coli
Neutrophils
Complementary DNA
Smoking
Lung
Inhibition (Psychology)

ASJC Scopus subject areas

  • Hematology

Cite this

Antiprotease targeting : altered specificity of α1-antitrypsin by amino acid replacement at the reactive centre. / Jallat, S.; Tessier, L. H.; Benavente, A.; Crystal, Ronald; Courtney, M.

In: Revue Francaise de Transfusion et Immuno-Hematologie, Vol. 29, No. 4, 01.01.1986, p. 287-298.

Research output: Contribution to journalArticle

@article{d96684b5a4ee4885866e955c79b6cb0b,
title = "Antiprotease targeting: altered specificity of α1-antitrypsin by amino acid replacement at the reactive centre",
abstract = "Alpha-1 antitrypsin (α1AT) is an efficient inhibitor of the human neutrophil proteases, elastase and cathepsin G. The reactive centre P1 residue (Met358) of α1AT is important in defining the specificity of inhibition; furthermore, oxidation of this residue results in a loss of inhibitor activity. There is evidence that oxidative inactivation of α1AT may be involved in the pathogenesis of pulmonary emphysema associated with cigarette smoking. We have studied the effect of a series of amino acid replacements at the active centre on the inhibition properties of α1AT. The mutant proteins were produced in E. coli following in vitro mutagenesis of the α1AT cDNA. Alpha-1-AT (Ile358), (Ala358) and (Val558) were efficient inhibitors of both neutrophil and pancreatic elastase, but not cathepsin G. Alpha-1-AT (Ala356, Val358) and α1AT (Phe358) were specific for pancreatic elastase and cathepsin G respectively. Alpha-1-AT (Leu358) inhibited both neutrophil elastase and cathepsin G. These data show that, for effective inhibition, a potential cleavage site for the protease must be displayed at the α1AT active centre. In each case, replacement of Met358 led to resistance to oxidative inactivation. Since α1AT (Leu358) inhibits both neutrophil proteases and is resistant to oxidation, this variant may be of increased potential for the therapy of destructive lung disorders.",
author = "S. Jallat and Tessier, {L. H.} and A. Benavente and Ronald Crystal and M. Courtney",
year = "1986",
month = "1",
day = "1",
doi = "10.1016/S0338-4535(86)80021-6",
language = "English",
volume = "29",
pages = "287--298",
journal = "Transfusion Clinique et Biologique",
issn = "1246-7820",
publisher = "Elsevier Masson",
number = "4",

}

TY - JOUR

T1 - Antiprotease targeting

T2 - altered specificity of α1-antitrypsin by amino acid replacement at the reactive centre

AU - Jallat, S.

AU - Tessier, L. H.

AU - Benavente, A.

AU - Crystal, Ronald

AU - Courtney, M.

PY - 1986/1/1

Y1 - 1986/1/1

N2 - Alpha-1 antitrypsin (α1AT) is an efficient inhibitor of the human neutrophil proteases, elastase and cathepsin G. The reactive centre P1 residue (Met358) of α1AT is important in defining the specificity of inhibition; furthermore, oxidation of this residue results in a loss of inhibitor activity. There is evidence that oxidative inactivation of α1AT may be involved in the pathogenesis of pulmonary emphysema associated with cigarette smoking. We have studied the effect of a series of amino acid replacements at the active centre on the inhibition properties of α1AT. The mutant proteins were produced in E. coli following in vitro mutagenesis of the α1AT cDNA. Alpha-1-AT (Ile358), (Ala358) and (Val558) were efficient inhibitors of both neutrophil and pancreatic elastase, but not cathepsin G. Alpha-1-AT (Ala356, Val358) and α1AT (Phe358) were specific for pancreatic elastase and cathepsin G respectively. Alpha-1-AT (Leu358) inhibited both neutrophil elastase and cathepsin G. These data show that, for effective inhibition, a potential cleavage site for the protease must be displayed at the α1AT active centre. In each case, replacement of Met358 led to resistance to oxidative inactivation. Since α1AT (Leu358) inhibits both neutrophil proteases and is resistant to oxidation, this variant may be of increased potential for the therapy of destructive lung disorders.

AB - Alpha-1 antitrypsin (α1AT) is an efficient inhibitor of the human neutrophil proteases, elastase and cathepsin G. The reactive centre P1 residue (Met358) of α1AT is important in defining the specificity of inhibition; furthermore, oxidation of this residue results in a loss of inhibitor activity. There is evidence that oxidative inactivation of α1AT may be involved in the pathogenesis of pulmonary emphysema associated with cigarette smoking. We have studied the effect of a series of amino acid replacements at the active centre on the inhibition properties of α1AT. The mutant proteins were produced in E. coli following in vitro mutagenesis of the α1AT cDNA. Alpha-1-AT (Ile358), (Ala358) and (Val558) were efficient inhibitors of both neutrophil and pancreatic elastase, but not cathepsin G. Alpha-1-AT (Ala356, Val358) and α1AT (Phe358) were specific for pancreatic elastase and cathepsin G respectively. Alpha-1-AT (Leu358) inhibited both neutrophil elastase and cathepsin G. These data show that, for effective inhibition, a potential cleavage site for the protease must be displayed at the α1AT active centre. In each case, replacement of Met358 led to resistance to oxidative inactivation. Since α1AT (Leu358) inhibits both neutrophil proteases and is resistant to oxidation, this variant may be of increased potential for the therapy of destructive lung disorders.

UR - http://www.scopus.com/inward/record.url?scp=0022784085&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022784085&partnerID=8YFLogxK

U2 - 10.1016/S0338-4535(86)80021-6

DO - 10.1016/S0338-4535(86)80021-6

M3 - Article

C2 - 3544150

AN - SCOPUS:0022784085

VL - 29

SP - 287

EP - 298

JO - Transfusion Clinique et Biologique

JF - Transfusion Clinique et Biologique

SN - 1246-7820

IS - 4

ER -