Proteolytic inactivation of alpha-1-anti-chymotrypsin. Sites of cleavage and generation of chemotactic activity.
The effect of several microbial and mammalian proteinases on the inhibitory activity of human plasma alpha-1-anti-chymotrypsin (alpha-1-Achy) has been tested. Most of these enzymes caused rapid inactivation of the inhibitor by cleavage at single sites within the reactive-site loop between P5 Lys and P3' Leu, with additional cleavages also being detected in some cases near the NH2 terminus of the native protein. In contrast, two of the enzymes tested failed to inactivate alpha-1-Achy, although they could cause removal of peptides near the NH2 terminus. Studies of neutrophil chemotaxis revealed that native or NH2-terminally truncated alpha-1-Achy was not stimulatory. However, testing of two enzymatically inactivated forms of the inhibitor (alpha-1-Achy), cleaved at widely different positions within the reactive-site loop, indicated that they had become potent chemoattractants at concentrations within the nanomolar range. Competition studies using proteolytically inactivated alpha-1-proteinase inhibitor suggested that the chemotactic activity of the two inactivated serpins was probably mediated by the same mechanism. The physiological relevance of this chemotactic activity is underscored by the results of plasma elimination studies which indicate that alpha-1-Achy is eliminated at approximately the same rate as native alpha-1-Achy, thus prolonging chemotactic stimuli within the tissues.
Potempa
J
J
Institute of Molecular Biology, Jagiellonian University, Cracow, Poland.
Fedak
D
D
Dubin
A
A
Mast
A
A
Travis
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008657:Metabolic Clearance Rate; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014357:Trypsin / Q000378:metabolism; D000514:alpha 1-Antichymotrypsin / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D000514:alpha 1-Antichymotrypsin; D010450:Endopeptidases; D014357:Trypsin