Catalytic activity and inhibition of wegener antigen proteinase 3 on the cell surface of human polymorphonuclear neutrophils.
Proteinase 3 (Pr3), the main target of anti-neutrophil cytoplasmic antibodies, is a neutrophil serine protease that may be constitutively expressed at the surface of quiescent circulating neutrophils. This raises the question of the simultaneous presence in the circulation of constitutive membrane-bound Pr3 (mPr3) and its plasma inhibitor alpha1-protease inhibitor (alpha1-Pi). We have looked at the fate of constitutive mPr3 at the surface of circulating blood neutrophils and of induced mPr3 on triggered neutrophils. We found significant Pr3 activity at the surface of activated neutrophils but not at the surface of quiescent neutrophils whatever the constitutive expression. This suggests that constitutive mPr3 is enzymatically inactive or its active site is not accessible to the substrate. Supporting this conclusion, we have not been able to demonstrate any interaction between constitutive mPr3 and alpha1-Pi, whereas induced mPr3 is cleared from the cell surface when activated cells are incubated with this inhibitor. But, unlike membrane-bound elastase that is also cleared from the surface of activated cells, mPr3 remained bound to the membrane when inhibited by elafin or by a low molecular weight chloromethyl ketone inhibitor, which shows that it binds more tightly to the neutrophil membrane. mPr3 may thus be present at the surface of circulating neutrophils in an environment replete with alpha1-Pi. The permanent presence of inactive Pr3 at the surface of quiescent neutrophils may explain why Pr3 is a major target of anti-neutrophil cytoplasmic antibodies, whose binding activates neutrophils and triggers inflammation, as in Wegener granulomatosis.
Korkmaz
Brice
B
INSERM U618, Protéases et Vectorisation Pulmonaires, Faculté de Médecine, 37032 Tours Cedex, France.
Jaillet
Jérôme
J
Jourdan
Marie-Lise
ML
Gauthier
Alexandre
A
Gauthier
Francis
F
Attucci
Sylvie
S
D000590:Amino Acid Chloromethyl Ketones / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D053492:Elafin / Q000378:metabolism; D004795:Enzyme Stability; D014890:Granulomatosis with Polyangiitis / Q000201:enzymology / Q000276:immunology; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism; D053146:Myeloblastin / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000201:enzymology* / Q000276:immunology*; D011485:Protein Binding; D000515:alpha 1-Antitrypsin / Q000378:metabolism*
D000590:Amino Acid Chloromethyl Ketones; D053492:Elafin; D000515:alpha 1-Antitrypsin; D019272:Leukocyte Elastase; D053146:Myeloblastin