The human airway trypsin-like protease modulates the urokinase receptor (uPAR, CD87) structure and functions.
The human airway trypsin-like protease (HAT) is a respiratory epithelium-associated, type II transmembrane serine protease, which is also detected as an extracellular enzyme in lung fluids during airway inflammatory disorders. We have evaluated its capacity to affect the urokinase-type plasminogen activator receptor (uPAR), a membrane glycolipid-anchored, three-domain (D1D2D3) glycoprotein that plays a crucial role in innate immunity and inflammation by supporting cell migration and matrix degradation, with structure and biological properties that can be regulated via limited endoproteolysis. With the use of immunoblotting, flow immunocytometry, and ELISA analyses applied to a recombinant uPAR protein and to uPAR-expressing monocytic and human bronchial epithelial cells, it was shown that exposure of uPAR to soluble HAT in the range of 10-500 nM resulted in the proteolytic processing of the full-length (D1D2D3) into the truncated (D2D3) species, with cleavage occurring in the D1 to D2 linker sequence after arginine residues at position 83 and 89. Using immunohistochemistry, we found that both HAT and uPAR were expressed in the human bronchial epithelium. Moreover, transient cotransfection in epithelial cells showed that membrane coexpression of the two partners produced a constitutive and extensive shedding of the D1 domain, occurring for membrane-associated HAT concentrations in the nanomolar range. Because the truncated receptor was found to be unable to bind two of the major uPAR ligands, the adhesive matrix protein vitronectin and the serine protease urokinase, it thus appears that proteolytic regulation of uPAR by HAT is likely to modulate cell adherence and motility, as well as tissue remodeling during the inflammatory response in the airways.
Beaufort
Nathalie
N
Inserm, E0336, Paris, France.
Leduc
Dominique
D
Eguchi
Hiroshi
H
Mengele
Karin
K
Hellmann
Daniela
D
Masegi
Tsukio
T
Kamimura
Takashi
T
Yasuoka
Susumu
S
Fend
Falko
F
Chignard
Michel
M
Pidard
Dominique
D
D002448:Cell Adhesion; D002465:Cell Movement; D017931:DNA Primers; D006801:Humans; D007249:Inflammation / Q000473:pathology / Q000503:physiopathology*; D008168:Lung / Q000473:pathology / Q000502:physiology / Q000503:physiopathology; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000378:metabolism; D020545:Respiratory Mucosa / Q000502:physiology*; D012697:Serine Endopeptidases / Q000378:metabolism*; D014746:Vimentin / Q000502:physiology
D017931:DNA Primers; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D014746:Vimentin; C114880:trypsin-like serine protease; D012697:Serine Endopeptidases