Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments

Olaf Wiesner, Robert D. Litwiller, Amber M. Hummel, Margaret A. Viss, Cari J. McDonald, Dieter E. Jenne, David N. Fass, Ulrich Specks

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Direct comparisons of human (h) and murine (m) neutrophil elastase (NE) and proteinase 3 (PR3) are important for the understanding and interpretation of inflammatory and PR3-related autoimmune processes investigated in wild-type-, mNE- and mPR3/mNE knockout mice. To this end, we purified recombinant mPR3 and mNE expressed in HMC1 and 293 cells and compared their biophysical properties, proteolytic activities and susceptibility to inhibitors with those of their human homologues, hPR3 and hNE. Significant species differences in physico-chemical properties, substrate specificities and enzyme kinetics towards synthetic peptide substrates, oxidized insulin B chain, and fibrinogen were detected. MeOSuc-AAPV-pNA and Suc-AAPV-pNA were hydrolyzed more efficiently by mPR3 than hPR3, but enzymatic activities of mNE and hNE were very similar. Fibrinogen was cleaved much more efficiently by mPR3 than by hPR3. All four proteases were inhibited by α1-antitrypsin and elafin. Eglin C inihibited mNE, hNE, mPR3, but not hPR3. SLPI inhibited both NEs, but neither PR3. The custom-designed hNE inhibitor, Val15-aprotinin, is a poor inhibitor for mNE. In conclusion, appropriate interpretation of experiments in murine models requires individual species-specific assessment of neutrophil protease function and inhibition.

Original languageEnglish (US)
Pages (from-to)5305-5312
Number of pages8
JournalFEBS Letters
Volume579
Issue number24
DOIs
StatePublished - Oct 10 2005

Fingerprint

Myeloblastin
Leukocyte Elastase
Fibrinogen
Elafin
Peptide Hydrolases
Enzyme kinetics
Aprotinin
Experiments
Substrates
Substrate Specificity
Knockout Mice
Chemical properties
Neutrophils
Insulin
Peptides
Enzymes

Keywords

  • ANCA
  • Neutrophil elastase
  • Neutrophil serine protease
  • Proteinase 3
  • Species differences

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments. / Wiesner, Olaf; Litwiller, Robert D.; Hummel, Amber M.; Viss, Margaret A.; McDonald, Cari J.; Jenne, Dieter E.; Fass, David N.; Specks, Ulrich.

In: FEBS Letters, Vol. 579, No. 24, 10.10.2005, p. 5305-5312.

Research output: Contribution to journalArticle

Wiesner, Olaf ; Litwiller, Robert D. ; Hummel, Amber M. ; Viss, Margaret A. ; McDonald, Cari J. ; Jenne, Dieter E. ; Fass, David N. ; Specks, Ulrich. / Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments. In: FEBS Letters. 2005 ; Vol. 579, No. 24. pp. 5305-5312.
@article{ec9ea0058cc14e0eaee10d8e83b5872d,
title = "Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments",
abstract = "Direct comparisons of human (h) and murine (m) neutrophil elastase (NE) and proteinase 3 (PR3) are important for the understanding and interpretation of inflammatory and PR3-related autoimmune processes investigated in wild-type-, mNE- and mPR3/mNE knockout mice. To this end, we purified recombinant mPR3 and mNE expressed in HMC1 and 293 cells and compared their biophysical properties, proteolytic activities and susceptibility to inhibitors with those of their human homologues, hPR3 and hNE. Significant species differences in physico-chemical properties, substrate specificities and enzyme kinetics towards synthetic peptide substrates, oxidized insulin B chain, and fibrinogen were detected. MeOSuc-AAPV-pNA and Suc-AAPV-pNA were hydrolyzed more efficiently by mPR3 than hPR3, but enzymatic activities of mNE and hNE were very similar. Fibrinogen was cleaved much more efficiently by mPR3 than by hPR3. All four proteases were inhibited by α1-antitrypsin and elafin. Eglin C inihibited mNE, hNE, mPR3, but not hPR3. SLPI inhibited both NEs, but neither PR3. The custom-designed hNE inhibitor, Val15-aprotinin, is a poor inhibitor for mNE. In conclusion, appropriate interpretation of experiments in murine models requires individual species-specific assessment of neutrophil protease function and inhibition.",
keywords = "ANCA, Neutrophil elastase, Neutrophil serine protease, Proteinase 3, Species differences",
author = "Olaf Wiesner and Litwiller, {Robert D.} and Hummel, {Amber M.} and Viss, {Margaret A.} and McDonald, {Cari J.} and Jenne, {Dieter E.} and Fass, {David N.} and Ulrich Specks",
year = "2005",
month = "10",
day = "10",
doi = "10.1016/j.febslet.2005.08.056",
language = "English (US)",
volume = "579",
pages = "5305--5312",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "24",

}

TY - JOUR

T1 - Differences between human proteinase 3 and neutrophil elastase and their murine homologues are relevant for murine model experiments

AU - Wiesner, Olaf

AU - Litwiller, Robert D.

AU - Hummel, Amber M.

AU - Viss, Margaret A.

AU - McDonald, Cari J.

AU - Jenne, Dieter E.

AU - Fass, David N.

AU - Specks, Ulrich

PY - 2005/10/10

Y1 - 2005/10/10

N2 - Direct comparisons of human (h) and murine (m) neutrophil elastase (NE) and proteinase 3 (PR3) are important for the understanding and interpretation of inflammatory and PR3-related autoimmune processes investigated in wild-type-, mNE- and mPR3/mNE knockout mice. To this end, we purified recombinant mPR3 and mNE expressed in HMC1 and 293 cells and compared their biophysical properties, proteolytic activities and susceptibility to inhibitors with those of their human homologues, hPR3 and hNE. Significant species differences in physico-chemical properties, substrate specificities and enzyme kinetics towards synthetic peptide substrates, oxidized insulin B chain, and fibrinogen were detected. MeOSuc-AAPV-pNA and Suc-AAPV-pNA were hydrolyzed more efficiently by mPR3 than hPR3, but enzymatic activities of mNE and hNE were very similar. Fibrinogen was cleaved much more efficiently by mPR3 than by hPR3. All four proteases were inhibited by α1-antitrypsin and elafin. Eglin C inihibited mNE, hNE, mPR3, but not hPR3. SLPI inhibited both NEs, but neither PR3. The custom-designed hNE inhibitor, Val15-aprotinin, is a poor inhibitor for mNE. In conclusion, appropriate interpretation of experiments in murine models requires individual species-specific assessment of neutrophil protease function and inhibition.

AB - Direct comparisons of human (h) and murine (m) neutrophil elastase (NE) and proteinase 3 (PR3) are important for the understanding and interpretation of inflammatory and PR3-related autoimmune processes investigated in wild-type-, mNE- and mPR3/mNE knockout mice. To this end, we purified recombinant mPR3 and mNE expressed in HMC1 and 293 cells and compared their biophysical properties, proteolytic activities and susceptibility to inhibitors with those of their human homologues, hPR3 and hNE. Significant species differences in physico-chemical properties, substrate specificities and enzyme kinetics towards synthetic peptide substrates, oxidized insulin B chain, and fibrinogen were detected. MeOSuc-AAPV-pNA and Suc-AAPV-pNA were hydrolyzed more efficiently by mPR3 than hPR3, but enzymatic activities of mNE and hNE were very similar. Fibrinogen was cleaved much more efficiently by mPR3 than by hPR3. All four proteases were inhibited by α1-antitrypsin and elafin. Eglin C inihibited mNE, hNE, mPR3, but not hPR3. SLPI inhibited both NEs, but neither PR3. The custom-designed hNE inhibitor, Val15-aprotinin, is a poor inhibitor for mNE. In conclusion, appropriate interpretation of experiments in murine models requires individual species-specific assessment of neutrophil protease function and inhibition.

KW - ANCA

KW - Neutrophil elastase

KW - Neutrophil serine protease

KW - Proteinase 3

KW - Species differences

UR - http://www.scopus.com/inward/record.url?scp=25844445300&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=25844445300&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2005.08.056

DO - 10.1016/j.febslet.2005.08.056

M3 - Article

C2 - 16182289

AN - SCOPUS:25844445300

VL - 579

SP - 5305

EP - 5312

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 24

ER -