Host defense functions of proteolytically processed and parent (unprocessed) cathelicidins of rabbit granulocytes.
Members of the cathelicidin family are present in all mammals studied. Generally, these proteins contain a conserved N-terminal domain and a structurally and functionally divergent C-terminal region that expresses antibacterial or other activities when proteolytically released. Rabbit granulocytes produce CAP18, a cathelicidin that conforms to this structural and functional organization, and also 15-kDa protein isoforms (p15s) that share several key structural features with other cathelicidins but apparently do not undergo processing with release of an active peptide. To further define the importance of proteolysis in the antibacterial activities of these proteins, we have purified from granulocytes proCAP18, its C-terminal peptide (CAP18p), and two p15 isoforms to apparent homogeneity. Of these four polypeptides, only CAP18p was independently cytotoxic to encapsulated Escherichia coli (90% inhibitory concentration, approximately 600 nM) but it was approximately 50-fold less potent on a molar basis than the bactericidal/permeability-increasing protein (BPI). However, all four cathelicidin species, notably including proCAP18, exhibited antibacterial synergy with BPI, and the p15s also displayed synergy with CAP18p in the absence of BPI. Subnanomolar concentrations of proCAP18 blocked lipopolysaccharide-induced chemiluminescence of human leukocytes, showing a molar potency more than 100-fold greater than that of CAP18p ( approximately 20 nM) or BPI ( approximately 50 nM). Thus, while independent bactericidal activity of cathelicidins requires processing, other host-defense functions do not and are more potently expressed by the unprocessed protein than by the C-terminal peptide.
Zarember
Kol A
KA
Department of Microbiology, School of Medicine New York University, New York, New York 10016, USA. kol@gene.com
Katz
Seth S
SS
Tack
Brian F
BF
Doukhan
Laurence
L
Weiss
Jerrold
J
Elsbach
Peter
P
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D001798:Blood Proteins / Q000494:pharmacology; D054804:Cathelicidins; D003470:Culture Media; D004336:Drug Antagonism; D004357:Drug Synergism; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects; D006801:Humans; D007552:Isotonic Solutions; D008070:Lipopolysaccharides / Q000494:pharmacology; D008565:Membrane Proteins; D009504:Neutrophils / Q000378:metabolism*; D020033:Protein Isoforms / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D011498:Protein Precursors / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D011499:Protein Processing, Post-Translational; D011817:Rabbits
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D054804:Cathelicidins; D003470:Culture Media; D007552:Isotonic Solutions; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D020033:Protein Isoforms; D011498:Protein Precursors; C021623:bactericidal permeability increasing protein; D010450:Endopeptidases