Inhibition of neutrophil elastase by recombinant human proteinase inhibitor 9.
Proteinase inhibitor PI9 (PI9) is an intracellular 42-kDa member of the ovalbumin family of serpins that is found primarily in placenta, lung and lymphocytes. PI9 has been shown to be a fast-acting inhibitor of granzyme B in vitro, presumably through the utilization of Glu(340) as the P(1) inhibitory residue in its reactive site loop. In this report, we describe the inhibition of human neutrophil elastase by recombinant human PI9. Inhibition occurred with an overall K(i)' of 221 pM and a second-order association rate constant of 1.5 x 10(5) M(-1) s(-1), indicating that PI9 is a potent inhibitor of this serine proteinase in vitro. In addition, incubation of recombinant PI9 with native neutrophil elastase resulted in the formation of an SDS-resistant 62-kDa complex. Amino-terminal sequence analyses provided evidence that inhibition of elastase occurred through the use of Cys(342) as the reactive P(1) amino acid residue in the PI9 reactive site loop. Thus, PI9 joins its close relatives PI6 and PI8 as having the ability to utilize multiple reactive site loop residues as the inhibitory P(1) residue to expand its inhibitory spectrum.
Dahlen
J R
JR
Department of Pathology, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA.
Foster
D C
DC
Kisiel
W
W
D001665:Binding Sites; D053804:Granzymes; D007700:Kinetics; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011506:Proteins / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000737:chemistry; D015843:Serpins / Q000737:chemistry / Q000494:pharmacology*
D011506:Proteins; D011994:Recombinant Proteins; C102316:SERPINB9 protein, human; D015843:Serpins; C075177:SERPINB1 protein, human; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases; D019272:Leukocyte Elastase