Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma

Melissa A. Fath, Xiaojun Wu, Ronald E. Hileman, Robert J. Linhardt, Mohammed A. Kashem, Richard M. Nelson, Clifford D. Wright, William M. Abraham

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Protease inhibition by secretory leukocyte protease inhibitor (SLPI) is accelerated by the sulfated polysaccharides. The nature of the SLPI- polysaccharide interaction, explored with affinity chromatography, indicated that this interaction was sensitive to the charge and type of polysaccharide. Dextran and chondroitin had the lowest affinity for SLPI, followed by dermatan, heparan, and dextran sulfates. While heparin bound SLPI tightly, the highest affinity heparin chains unexpectedly contained a lower level of sulfation than more weakly interacting chains. Heparin oligosaccharides, prepared using heparin lyase I were SLPI-affinity fractionated. Surprisingly, undersulfated heparin oligosaccharides bound SLPI with the highest affinity, suggesting the importance of free hydroxyl groups for high affinity interaction. Isothermal titration calorimetry was used to determine the thermodynamics of SLPI interaction with a low molecular weight heparin, an undersulfated decasaccharide and a tetrasaccharide. The studies showed 12-14 saccharide units, corresponding to molecular weight of ~4,800, were required for a 1:1 (SLPI:heparin) binding stoichiometry. Furthermore, an undersulfated decasaccharide was able to bind SLPI tightly (K(d) ~13 nM), resulting in its activation and the inhibition of neutrophil elastase and pancreatic chymotrypsin. The in vitro assessment of heparin and the decasaccharide and tetrasaccharide using stopped-flow kinetics suggested that heparin was the optimal choice to study SLPI-based in vivo protease inhibition. SLPI and heparin were co-administered by inhalation in therapy against antigen- induced airway hyperresponsiveness in a sheep bronchoprovocation model. Heparin, in combination with SLPI demonstrated in vivo efficacy reducing early and late phase bronchoconstriction. Heparin also increased the therapeutic activity of SLPI against antigen-induced airway hyperresponsiveness.

Original languageEnglish
Pages (from-to)13563-13569
Number of pages7
JournalJournal of Biological Chemistry
Volume273
Issue number22
DOIs
StatePublished - May 29 1998
Externally publishedYes

Fingerprint

Secretory Leukocyte Peptidase Inhibitor
Heparin
Peptide Hydrolases
Asthma
Polysaccharides
Oligosaccharides
Chondroitin
Heparin Lyase
Affinity chromatography
Respiratory Therapy
Antigens
Dermatan Sulfate
Leukocyte Elastase
Dextran Sulfate
Calorimetry
Bronchoconstriction
Heparitin Sulfate
Low Molecular Weight Heparin
Chymotrypsin
Dextrans

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fath, M. A., Wu, X., Hileman, R. E., Linhardt, R. J., Kashem, M. A., Nelson, R. M., ... Abraham, W. M. (1998). Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma. Journal of Biological Chemistry, 273(22), 13563-13569. https://doi.org/10.1074/jbc.273.22.13563

Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma. / Fath, Melissa A.; Wu, Xiaojun; Hileman, Ronald E.; Linhardt, Robert J.; Kashem, Mohammed A.; Nelson, Richard M.; Wright, Clifford D.; Abraham, William M.

In: Journal of Biological Chemistry, Vol. 273, No. 22, 29.05.1998, p. 13563-13569.

Research output: Contribution to journalArticle

Fath, MA, Wu, X, Hileman, RE, Linhardt, RJ, Kashem, MA, Nelson, RM, Wright, CD & Abraham, WM 1998, 'Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma', Journal of Biological Chemistry, vol. 273, no. 22, pp. 13563-13569. https://doi.org/10.1074/jbc.273.22.13563
Fath, Melissa A. ; Wu, Xiaojun ; Hileman, Ronald E. ; Linhardt, Robert J. ; Kashem, Mohammed A. ; Nelson, Richard M. ; Wright, Clifford D. ; Abraham, William M. / Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 22. pp. 13563-13569.
@article{a01c19e833f44670acd7bf506c598f70,
title = "Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma",
abstract = "Protease inhibition by secretory leukocyte protease inhibitor (SLPI) is accelerated by the sulfated polysaccharides. The nature of the SLPI- polysaccharide interaction, explored with affinity chromatography, indicated that this interaction was sensitive to the charge and type of polysaccharide. Dextran and chondroitin had the lowest affinity for SLPI, followed by dermatan, heparan, and dextran sulfates. While heparin bound SLPI tightly, the highest affinity heparin chains unexpectedly contained a lower level of sulfation than more weakly interacting chains. Heparin oligosaccharides, prepared using heparin lyase I were SLPI-affinity fractionated. Surprisingly, undersulfated heparin oligosaccharides bound SLPI with the highest affinity, suggesting the importance of free hydroxyl groups for high affinity interaction. Isothermal titration calorimetry was used to determine the thermodynamics of SLPI interaction with a low molecular weight heparin, an undersulfated decasaccharide and a tetrasaccharide. The studies showed 12-14 saccharide units, corresponding to molecular weight of ~4,800, were required for a 1:1 (SLPI:heparin) binding stoichiometry. Furthermore, an undersulfated decasaccharide was able to bind SLPI tightly (K(d) ~13 nM), resulting in its activation and the inhibition of neutrophil elastase and pancreatic chymotrypsin. The in vitro assessment of heparin and the decasaccharide and tetrasaccharide using stopped-flow kinetics suggested that heparin was the optimal choice to study SLPI-based in vivo protease inhibition. SLPI and heparin were co-administered by inhalation in therapy against antigen- induced airway hyperresponsiveness in a sheep bronchoprovocation model. Heparin, in combination with SLPI demonstrated in vivo efficacy reducing early and late phase bronchoconstriction. Heparin also increased the therapeutic activity of SLPI against antigen-induced airway hyperresponsiveness.",
author = "Fath, {Melissa A.} and Xiaojun Wu and Hileman, {Ronald E.} and Linhardt, {Robert J.} and Kashem, {Mohammed A.} and Nelson, {Richard M.} and Wright, {Clifford D.} and Abraham, {William M.}",
year = "1998",
month = "5",
day = "29",
doi = "10.1074/jbc.273.22.13563",
language = "English",
volume = "273",
pages = "13563--13569",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "22",

}

TY - JOUR

T1 - Interaction of secretory leukocyte protease inhibitor with heparin inhibits proteases involved in asthma

AU - Fath, Melissa A.

AU - Wu, Xiaojun

AU - Hileman, Ronald E.

AU - Linhardt, Robert J.

AU - Kashem, Mohammed A.

AU - Nelson, Richard M.

AU - Wright, Clifford D.

AU - Abraham, William M.

PY - 1998/5/29

Y1 - 1998/5/29

N2 - Protease inhibition by secretory leukocyte protease inhibitor (SLPI) is accelerated by the sulfated polysaccharides. The nature of the SLPI- polysaccharide interaction, explored with affinity chromatography, indicated that this interaction was sensitive to the charge and type of polysaccharide. Dextran and chondroitin had the lowest affinity for SLPI, followed by dermatan, heparan, and dextran sulfates. While heparin bound SLPI tightly, the highest affinity heparin chains unexpectedly contained a lower level of sulfation than more weakly interacting chains. Heparin oligosaccharides, prepared using heparin lyase I were SLPI-affinity fractionated. Surprisingly, undersulfated heparin oligosaccharides bound SLPI with the highest affinity, suggesting the importance of free hydroxyl groups for high affinity interaction. Isothermal titration calorimetry was used to determine the thermodynamics of SLPI interaction with a low molecular weight heparin, an undersulfated decasaccharide and a tetrasaccharide. The studies showed 12-14 saccharide units, corresponding to molecular weight of ~4,800, were required for a 1:1 (SLPI:heparin) binding stoichiometry. Furthermore, an undersulfated decasaccharide was able to bind SLPI tightly (K(d) ~13 nM), resulting in its activation and the inhibition of neutrophil elastase and pancreatic chymotrypsin. The in vitro assessment of heparin and the decasaccharide and tetrasaccharide using stopped-flow kinetics suggested that heparin was the optimal choice to study SLPI-based in vivo protease inhibition. SLPI and heparin were co-administered by inhalation in therapy against antigen- induced airway hyperresponsiveness in a sheep bronchoprovocation model. Heparin, in combination with SLPI demonstrated in vivo efficacy reducing early and late phase bronchoconstriction. Heparin also increased the therapeutic activity of SLPI against antigen-induced airway hyperresponsiveness.

AB - Protease inhibition by secretory leukocyte protease inhibitor (SLPI) is accelerated by the sulfated polysaccharides. The nature of the SLPI- polysaccharide interaction, explored with affinity chromatography, indicated that this interaction was sensitive to the charge and type of polysaccharide. Dextran and chondroitin had the lowest affinity for SLPI, followed by dermatan, heparan, and dextran sulfates. While heparin bound SLPI tightly, the highest affinity heparin chains unexpectedly contained a lower level of sulfation than more weakly interacting chains. Heparin oligosaccharides, prepared using heparin lyase I were SLPI-affinity fractionated. Surprisingly, undersulfated heparin oligosaccharides bound SLPI with the highest affinity, suggesting the importance of free hydroxyl groups for high affinity interaction. Isothermal titration calorimetry was used to determine the thermodynamics of SLPI interaction with a low molecular weight heparin, an undersulfated decasaccharide and a tetrasaccharide. The studies showed 12-14 saccharide units, corresponding to molecular weight of ~4,800, were required for a 1:1 (SLPI:heparin) binding stoichiometry. Furthermore, an undersulfated decasaccharide was able to bind SLPI tightly (K(d) ~13 nM), resulting in its activation and the inhibition of neutrophil elastase and pancreatic chymotrypsin. The in vitro assessment of heparin and the decasaccharide and tetrasaccharide using stopped-flow kinetics suggested that heparin was the optimal choice to study SLPI-based in vivo protease inhibition. SLPI and heparin were co-administered by inhalation in therapy against antigen- induced airway hyperresponsiveness in a sheep bronchoprovocation model. Heparin, in combination with SLPI demonstrated in vivo efficacy reducing early and late phase bronchoconstriction. Heparin also increased the therapeutic activity of SLPI against antigen-induced airway hyperresponsiveness.

UR - http://www.scopus.com/inward/record.url?scp=0032577686&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032577686&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.22.13563

DO - 10.1074/jbc.273.22.13563

M3 - Article

C2 - 9593692

AN - SCOPUS:0032577686

VL - 273

SP - 13563

EP - 13569

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 22

ER -