Neutrophil-derived proteinase 3 induces kallikrein-independent release of a novel vasoactive kinin.
The kinin-forming pathway is activated on endothelial cells and neutrophils when high-molecular weight kininogen (HK) is cleaved by plasma kallikrein liberating bradykinin, a potent mediator of inflammation. Kinins are released during inflammatory conditions such as vasculitis, associated with neutrophil influx around blood vessels. Some patients with vasculitis have elevated plasma levels of neutrophil-derived proteinase 3 (PR3) and anti-PR3 Abs. This study investigated if neutrophil-derived PR3 could induce activation of the kinin pathway. PR3 incubated with HK, or a synthetic peptide derived from HK, induced breakdown and release of a novel tridecapeptide termed PR3-kinin, NH(2)-MKRPPGFSPFRSS-COOH, consisting of bradykinin with two additional amino acids on each terminus. The reaction was specific and inhibited by anti-PR3 and alpha(1)-antitrypsin. Recombinant wild-type PR3 incubated with HK induced HK breakdown, whereas mutated PR3, lacking enzymatic activity, did not. PR3-kinin bound to and activated human kinin B(1) receptors, but did not bind to B(2) receptors, expressed by transfected HEK293 cells in vitro. In human plasma PR3-kinin was further processed to the B(2) receptor agonist bradykinin. PR3-kinin exerted a hypotensive effect in vivo through both B(1) and B(2) receptors as demonstrated using wild-type and B(1) overexpressing rats as well as wild-type and B(2) receptor knockout mice. Neutrophil extracts from vasculitis patients and healthy controls contained comparable amounts of PR3 and induced HK proteolysis, an effect that was abolished when PR3 was immunoadsorbed. Neutrophil-derived PR3 can proteolyze HK and liberate PR3-kinin, thereby initiating kallikrein-independent activation of the kinin pathway.
Kahn
Robin
R
Department of Pediatrics, Clinical Sciences Lund, Lund University, Lund, Sweden.
Hellmark
Thomas
T
Leeb-Lundberg
L M Fredrik
LM
Akbari
Nasrin
N
Todiras
Mihail
M
Olofsson
Tor
T
Wieslander
Jörgen
J
Christensson
Anders
A
Westman
Kerstin
K
Bader
Michael
M
Müller-Esterl
Werner
W
Karpman
Diana
D
D000293:Adolescent; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001920:Bradykinin / Q000097:blood; D002648:Child; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D007704:Kininogens / Q000378:metabolism; D007705:Kinins / Q000378:metabolism*; D008297:Male; D051379:Mice; D008875:Middle Aged; D053146:Myeloblastin / Q000235:genetics / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology*; D011485:Protein Binding; D051381:Rats
D007704:Kininogens; D007705:Kinins; D007610:Kallikreins; D053146:Myeloblastin; D001920:Bradykinin