Attenuated kinin release from human neutrophil elastase-pretreated kininogens by tissue and plasma kallikreins.
Components of kinin-forming systems operating at inflammatory sites are likely to interact with elastase that is released by recruited neutrophils and may, at least temporarily, constitute the major proteolytic activity present at these sites. The aim of this work was to determine the effect of kininogen degradation by human neutrophil elastase (HNE) on kinin generation by tissue and plasma kallikreins. We show that the digestion of both low molecular mass (LK) and high molecular mass (HK) forms of human kininogen by HNE renders them essentially unsusceptible to processing by human urinary kallikrein (tissue-type) and also significantly quenches the kinin release from HK by plasma kallikrein. Studies with synthetic model heptadecapeptide substrates, ISLMKRPPGFSPFRSSR and SLMKRPPGFSPFRSSRI, confirmed the inability of tissue kallikrein to process peptides at either termini of the internal kinin sequence, while plasma kallikrein was shown to release the kinin C-terminus relatively easily. The HNE-generated fragments of kininogens were separated by HPLC and the fractions containing internal kinin sequences were identified by a kinin-specific immunoenzymatic test after trypsin digestion. These fractions were analyzed by electrospray-ionization mass spectrometry. In this way, multiple peptides containing the kinin sequence flanked by only a few amino acid residues at each terminus were identified in elastase digests of both LK and HK. These results suggest that elastase may be involved in quenching the kinin-release cascade at the late stages of the inflammatory reaction.
Dulinski
Robert
R
Faculty of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland.
Suder
Piotr
P
Guevara-Lora
Ibeth
I
Rapała-Kozik
Maria
M
Potempa
Jan
J
Silberring
Jerzy
J
Imamura
Takahisa
T
Travis
James
J
Kozik
Andrzej
A
D000595:Amino Acid Sequence; D006801:Humans; D007704:Kininogens / Q000737:chemistry / Q000378:metabolism*; D007705:Kinins / Q000737:chemistry / Q000378:metabolism*; D019272:Leukocyte Elastase / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D020842:Plasma Kallikrein / Q000378:metabolism*; D020840:Tissue Kallikreins / Q000378:metabolism*
D007704:Kininogens; D007705:Kinins; D010446:Peptide Fragments; D020842:Plasma Kallikrein; D020840:Tissue Kallikreins; D019272:Leukocyte Elastase