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 language | English (US) |
|---|---|
| Pages (from-to) | 5305-5312 |
| Number of pages | 8 |
| Journal | FEBS Letters |
| Volume | 579 |
| Issue number | 24 |
| DOIs | |
| State | Published - Oct 10 2005 |
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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 journal › Article
}
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 -