Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3.
Cathelicidins are a family of antimicrobial proteins found in the peroxidase-negative granules of neutrophils. The known biologic functions reside in the C-terminus, which must be cleaved from the holoprotein to become active. Bovine and porcine cathelicidins are cleaved by elastase from the azurophil granules to yield the active antimicrobial peptides. The aim of this study was to identify the physiological setting for cleavage of the only human cathelicidin, hCAP-18, to liberate the antibacterial and cytotoxic peptide LL-37 and to identify the protease responsible for this cleavage. Immunoelectron microscopy demonstrated that both hCAP-18 and azurophil granule proteins were present in the phagolysosome. Immunoblotting revealed no detectable cleavage of hCAP-18 in cells after phagocytosis. In contrast, hCAP-18 was cleaved to generate LL-37 in exocytosed material. Of the 3 known serine proteases from azurophil granules, proteinase 3 was solely responsible for cleavage of hCAP-18 after exocytosis. This is the first detailed study describing the generation of a human antimicrobial peptide from a promicrobicidal protein, and it demonstrates that the generation of active antimicrobial peptides from common proproteins occurs differently in related species. (Blood. 2001;97:3951-3959)
Sørensen
O E
OE
Granulocyte Research Laboratory, Department of Hematology, Copenhagen University Hospital, 9 Blegdamsvej, DK-2100 Copenhagen, Denmark. olesoeren@rh.dk
Follin
P
P
Johnsen
A H
AH
Calafat
J
J
Tjabringa
G S
GS
Hiemstra
P S
PS
Borregaard
N
N
D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D054804:Cathelicidins; D005089:Exocytosis; D006801:Humans; D015151:Immunoblotting; D008854:Microscopy, Electron; D053146:Myeloblastin; D009504:Neutrophils / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D010587:Phagocytosis; D012697:Serine Endopeptidases / Q000494:pharmacology*
D023181:Antimicrobial Cationic Peptides; D054804:Cathelicidins; D012697:Serine Endopeptidases; D053146:Myeloblastin