Alpha-defensins in enteric innate immunity: functional Paneth cell alpha-defensins in mouse colonic lumen.
Paneth cells are a secretory epithelial lineage that release dense core granules rich in host defense peptides and proteins from the base of small intestinal crypts. Enteric alpha-defensins, termed cryptdins (Crps) in mice, are highly abundant in Paneth cell secretions and inherently resistant to proteolysis. Accordingly, we tested the hypothesis that enteric alpha-defensins of Paneth cell origin persist in a functional state in the mouse large bowel lumen. To test this idea, putative Crps purified from mouse distal colonic lumen were characterized biochemically and assayed in vitro for bactericidal peptide activities. The peptides comigrated with cryptdin control peptides in acid-urea-PAGE and SDS-PAGE, providing identification as putative Crps. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry experiments showed that the molecular masses of the putative alpha-defensins matched those of the six most abundant known Crps, as well as N-terminally truncated forms of each, and that the peptides contain six Cys residues, consistent with identities as alpha-defensins. N-terminal sequencing definitively revealed peptides with N termini corresponding to full-length, (des-Leu)-truncated, and (des-Leu-Arg)-truncated N termini of Crps 1-4 and 6. Crps from mouse large bowel lumen were bactericidal in the low micromolar range. Thus, Paneth cell alpha-defensins secreted into the small intestinal lumen persist as intact and functional forms throughout the intestinal tract, suggesting that the peptides may mediate enteric innate immunity in the colonic lumen, far from their upstream point of secretion in small intestinal crypts.
Mastroianni
Jennifer R
JR
Department of Pathology and Laboratory Medicine, University of California, Irvine, California 92697-4800.
Ouellette
André J
AJ
Department of Pathology and Laboratory Medicine, University of California, Irvine, California 92697-4800; Department of Microbiology and Molecular Genetics, University of California, Irvine, California 92697-4800. Electronic address: andre.ouellette@keck.usc.edu.
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000276:immunology; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000276:immunology; D003106:Colon / Q000166:cytology / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology*; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D019879:Paneth Cells / Q000166:cytology / Q000276:immunology*; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology*; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D023084:alpha-Defensins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D023083:beta-Defensins / Q000235:genetics / Q000276:immunology
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D020033:Protein Isoforms; D011498:Protein Precursors; D023084:alpha-Defensins; D023083:beta-Defensins; C059569:cryptdin
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