Human leukocyte elastase is an endogenous ligand for the integrin CR3 (CD11b/CD18, Mac-1, alpha M beta 2) and modulates polymorphonuclear leukocyte adhesion.
Integrin CR3 (CD11b/CD18, Mac-1, alpha M beta 2) mediates the transient adhesion of polymorphonuclear leukocytes (PMN) to surfaces coated with fibrinogen, C3bi, ICAM-1, and other ligands. Recent studies (Cai, T.-Q., and S.D. Wright 1995. J. Biol. Chem. 270:14358) suggest that adhesion may be favored by stimulus-dependent changes in the kinetics of ligand binding by CR3. Cell detachment, on the other hand, must occur by a different mechanism because binding kinetics cannot affect cell adhesion after binding of ligand has occurred. We have sought a mechanism that would reverse binding of ligand to CR3 and report here that lysates of PMN contain an endogenous ligand that binds CR3 and competes the binding of C3bi. Purification and sequence analysis identified the structurally homologous azurophilic granule proteins, elastase, protease 3, and azurocidin as candidates. Studies with purified elastase and azurocidin showed that each bound specifically to purified, immobilized CR3. Elastase may play a role in modulating integrin-mediated cell adhesion because it is expressed at the cell surface, and the expression level is inversely proportional to cell adhesivity. Furthermore, a monoclonal antibody against elastase prevented detachment of PMN from fibrinogen-coated surfaces and blocked chemotaxis, confirming a role for this protein in regulating integrin-mediated adhesion. These studies suggest a model for release of integrin-mediated cell adhesion in which endogenous ligands such as elastase may release adhesion by "'eluting" substrate-bound ligand. A role for the proteolytic activity of elastase appears likely but is not demonstrated in this study.
Cai
T Q
TQ
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York 10021, USA.
Wright
S D
SD
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides; D001667:Binding, Competitive; D001798:Blood Proteins / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism*; D002352:Carrier Proteins; D002448:Cell Adhesion / Q000502:physiology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003179:Complement C3b / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D019272:Leukocyte Elastase / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008024:Ligands; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology*; D011480:Protease Inhibitors / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects
C062991:AZU1 protein, human; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D018821:CD18 Antigens; D002352:Carrier Proteins; D008024:Ligands; D016177:Macrophage-1 Antigen; D011480:Protease Inhibitors; D003179:Complement C3b; D005340:Fibrinogen; D019272:Leukocyte Elastase